本文由 上?；鈱崢I(yè)有限公司 整理匯編

2013-12-06 00:00 155閱讀次數(shù)

收藏 評價 舉報

• 分享

立即下載

參與評論

評論

登錄后參與評論

登錄或新用戶注冊

• 微信登錄
• 密碼登錄
• 短信登錄

請用手機微信掃描下方二維碼
快速登錄或注冊新賬號

微信掃碼，手機電腦聯(lián)動

注冊登錄即表示同意《儀器網(wǎng)服務(wù)條款》和《隱私協(xié)議》

賬  號：

密  碼：

圖片碼： 看不清？
換一張？

手機和郵箱號注冊新賬號>> 忘記密碼？

手機號：

圖片碼： 看不清？
換一張？

短信碼： 獲取短信碼

手機和郵箱號注冊新賬號>> 忘記密碼？

更多資料

• 人尿激酶(UK)ELISA試劑盒

  人尿激酶(UK)ELISA試劑盒[詳細]

  2013-12-06 00:00

  選購指南

  |

  • 
  • 
  • 
  • 

• 大鼠尿激酶(UK)ELISA試劑盒

  大鼠尿激酶(UK)ELISA試劑盒[詳細]

  2013-12-09 00:00

  報價單

  |

  • 
  • 
  • 
  • 

• 大鼠尿激酶（UK）ELISA試劑盒使用說明書

  大鼠尿激酶（UK）ELISA試劑盒使用說明書Elisakit規(guī)格：48孔配置/96孔配置標準品稀釋液：1.5ml×1瓶酶標試劑：3ml×1瓶（48）/6ml×1瓶（96）【大鼠尿激酶（UK）ELISA試劑盒】本試劑僅供研究使用計算：以標準物的濃度為橫坐標，OD值為縱坐標，在坐標紙上繪出標準曲線，根據(jù)樣品的OD值由標準曲線查出相應(yīng)的濃度；再乘以稀釋倍數(shù)；或用標準物的濃度與OD值計算出標準曲線的直線回歸方程式，將樣品的OD值代入方程式，計算出樣品濃度，再乘以稀釋倍數(shù)，即為樣品的實際濃度。試劑盒組成：封板膜:2片（48）/2片（96）說明書:1份密封袋:1個標準品：2700ng/L0.5ml×1瓶0.5ml×1瓶2-8℃保存酶標包被板:1×481×962-8℃保存樣品稀釋液:3ml×1瓶6ml×1瓶2-8℃保存顯色劑A液:3ml×1瓶6ml×1瓶2-8℃保存顯色劑B液:3ml×1瓶6ml×1瓶2-8℃保存終止液:3ml×1瓶6ml×1瓶2-8℃保存濃縮洗滌液:（20ml×20倍）×1瓶（20ml×30倍）×1瓶2-8℃保存實驗原理：本試劑盒應(yīng)用雙抗體夾心法測定標本中大鼠血血管生成素2（ANG-2）水平。用純化的大鼠血血管生成素2（ANG-2）抗體包被微孔板，制成固相抗體，往包被單抗的微孔中依次加入抗血小板抗體IgG/M/A（PA-IgG/M/A），再與HRP標記的抗血小板抗體IgG/M/A（PA-IgG/M/A）抗體結(jié)合，形成抗體-抗原-酶標抗體復(fù)合物，經(jīng)過徹底洗滌后加底物TMB顯色。TMB在HRP酶的催化下轉(zhuǎn)化成藍色，并在酸的作用下轉(zhuǎn)化成Z終的黃色。顏色的深淺和樣品中的抗血小板抗體IgG/M/A（PA-IgG/M/A）呈正相關(guān)。用酶標儀在450nm波長下測定吸光度（OD值），通過標準曲線計算樣品中大鼠血血管生成素2（ANG-2）濃度。目的：本試劑盒用于測大鼠血血清，血漿及相關(guān)液體樣本中抗血小板抗體IgG/M/A（PA-IgG/M/A）的含量。服務(wù)承諾：供貨期：款到發(fā)貨。工作時間內(nèi)免費的技術(shù)咨詢和指導(dǎo)。為客戶提供來樣檢測服務(wù)，Zda限度實驗結(jié)果的有效性（免費代測）。保存條件及有效期：試劑盒保存：2-8℃|有效期：6個月【大鼠尿激酶（UK）ELISA試劑盒】樣本處理及要求：1.血清：室溫血液自然凝固10-20分鐘，離心20分鐘左右（2000-3000轉(zhuǎn)/分）。仔細收集上清，保存過程中如出現(xiàn)沉淀，應(yīng)再次離心。2.血漿：應(yīng)根據(jù)標本的要求選擇EDTA或檸檬酸鈉作為抗凝劑，混合10-20分鐘后，離心20分鐘左右（2000-3000轉(zhuǎn)/分）。仔細收集上清，保存過程中如有沉淀形成，應(yīng)該再次離心。3.尿液：用無菌管收集，離心20分鐘左右（2000-3000轉(zhuǎn)/分）。仔細收集上清，保存過程中如有沉淀形成，應(yīng)再次離心。胸腹水、腦脊液參照實行。4.細胞培養(yǎng)上清：檢測分泌性的成份時，用無菌管收集。離心20分鐘左右（2000-3000轉(zhuǎn)/分）。仔細收集上清。檢測細胞內(nèi)的成份時，用PBS（PH7.2-7.4）稀釋細胞懸液，細胞濃度達到100萬/ml左右。通過反復(fù)凍融，以使細胞破壞并放出細胞內(nèi)成份。離心20分鐘左右（2000-3000轉(zhuǎn)/分）。仔細收集上清。保存過程中如有沉淀形成，應(yīng)再次離心。5.組織標本：切割標本后，稱取重量。加入一定量的PBS，PH7.4。用液氮迅速冷凍保存?zhèn)溆?。標本融化后仍然保?-8℃的溫度。加入一定量的PBS（PH7.4），用手工或勻漿器將標本勻漿充分。離心20分鐘左右（2000-3000轉(zhuǎn)/分）。仔細收集上清。分裝后一份待檢測，其余冷凍備用。6.標本采集后盡早進行提取，提取按相關(guān)文獻進行，提取后應(yīng)盡快進行實驗。若不能馬上進行試驗，可將標本放于-20℃保存，但應(yīng)避免反復(fù)凍融.7.不能檢測含NaN3的樣品，因NaN3YZ辣根過氧化物酶的（HRP）活性。操作步驟1.標準品的稀釋與加樣：在酶標包被板上設(shè)標準品孔10孔，在**、第二孔中分別加標準品100μl，然后在**、第二孔中加標準品稀釋液50μl，混勻；然后從**孔、第二孔中各取100μl分別加到第三孔和第四孔，再在第三、第四孔分別加標準品稀釋液50μl，混勻；然后在第三孔和第四孔中先各取50μl棄掉，再各取50μl分別加到第五、第六孔中，再在第五、第六孔中分別加標準品稀釋液50ul，混勻；混勻后從第五、第六孔中各取50μl分別加到第七、第八孔中，再在第七、第八孔中分別加標準品稀釋液50μl，混勻后從第七、第八孔中分別取50μl加到第九、第十孔中，再在第九第十孔分別加標準品稀釋液50μl，混勻后從第九第十孔中各取50μl棄掉。（稀釋后各孔加樣量都為50μl，濃度分別為1800ng/L，1200ng/L，600ng/L，300ng/L，150ng/L）。2.加樣：分別設(shè)空白孔（空白對照孔不加樣品及酶標試劑，其余各步操作相同）、待測樣品孔。在酶標包被板上待測樣品孔中先加樣品稀釋液40μl，然后再加待測樣品10μl（樣品Z終稀釋度為5倍）。加樣將樣品加于酶標板孔底部，盡量不觸及孔壁，輕輕晃動混勻。3.溫育：用封板膜封板后置37℃溫育30分鐘。4.配液：將30（48T的20倍）倍濃縮洗滌液用蒸餾水30（48T的20倍）倍稀釋后備用。5.洗滌：小心揭掉封板膜，棄去液體，甩干，每孔加滿洗滌液，靜置30秒后棄去，如此重復(fù)5次，拍干。6.加酶：每孔加入酶標試劑50μl，空白孔除外。7.溫育：操作同3。8.洗滌：操作同5。9.顯色：每孔先加入顯色劑A50μl，再加入顯色劑B50μl，輕輕震蕩混勻，37℃避光顯色15分鐘.10.終止：每孔加終止液50μl，終止反應(yīng)（此時藍色立轉(zhuǎn)黃色）。11.測定：以空白空調(diào)零，450nm波長依序測量各孔的吸光度（OD值）。測定應(yīng)在加終止液后15分鐘以內(nèi)進行?！敬笫竽蚣っ福║K）ELISA試劑盒】注意事項：1．試劑盒從冷藏環(huán)境中取出應(yīng)在室溫平衡15-30分鐘后方可使用，酶標包被板開封后如未用完，板條應(yīng)裝入密封袋中保存。2．濃洗滌液可能會有結(jié)晶析出，稀釋時可在水浴中加溫助溶，洗滌時不影響結(jié)果。3．各步加樣均應(yīng)使用加樣器，并經(jīng)常校對其準確性，以避免試驗誤差。一次加樣時間**控制在5分鐘內(nèi)，如標本數(shù)量多，推薦使用排槍加樣。4．請每次測定的同時做標準曲線，**做復(fù)孔。如標本中待測物質(zhì)含量過高（樣本OD值大于標準品孔**孔的OD值），請先用樣品稀釋液稀釋一定倍數(shù)（n倍）后再測定，計算時請Z后乘以總稀釋倍數(shù)（×n×5）。5．封板膜只限一次性使用，以避免交叉污染。6．底物請避光保存。7．嚴格按照說明書的操作進行，試驗結(jié)果判定必須以酶標儀讀數(shù)為準.8．所有樣品，洗滌液和各種廢棄物都應(yīng)按傳染物處理。9．本試劑不同批號組分不得混用。10.如與英文說明書有異，以英文說明書為準。試劑盒性能：1.樣品線性回歸與預(yù)期濃度相關(guān)系數(shù)R值為0.95以上。2.批內(nèi)與批見應(yīng)分別小于9%和11%檢測范圍：0.2IU/L-6IU/[詳細]

  2018-09-14 10:01

  產(chǎn)品樣冊

  |

  • 
  • 
  • 
  • 

• 人尿激酶型纖溶酶原激活物受體（PLAUR/uPAR）Elisa試劑盒說明書

  FORINVITROUSEANDRESEARCHUSEONLYNOTFORUSEINDIAGNOSTICORTHERAPEUTICPROCEDURES7thEdition（RevisedinNovember,2011）[INTENDEDUSE]ThekitisasandwichenzymeimmunoassayforinvitroquantitativemeasurementofuPARinhumanserum,plasmaandotherbiologicalfluids.[REAGENTSANDMATERIALSPROVIDED]ReagentsQuantityReagentsQuantityPre-coated,readytouse96-wellstripplate1Platesealerfor96wells4Standard（lyophilized）2StandardDiluent1×20mLDetectionReagentA（green）1×120μLAssayDiluentA（2×concentrate）1×6mLDetectionReagentB（red）1×120μLAssayDiluentB（2×concentrate）1×6mLTMBSubstrate1×9mLStopSolution1×6mLWashBuffer（30×concentrate）1×20mLInstructionmanual1[MATERIALSREQUIREDBUTNOTSUPPLIED]1.Microplatereaderwith450±10nmfilter.2.Precisionsingleormulti-channelpipettesanddisposabletips.3.EppendorfTubesfordilutingsamples.4.Deionizedordistilledwater.5.Absorbentpaperforblottingthemicrotiterplate.6.ContainerforWashSolution[STORAGEOFTHEKITS]1.Forunopenedkit:Allthereagents首ldbekeptaccordingtothelabelsonvials.TheStandard,DetectionReagentA,DetectionReagentBandthe96-wellstripplate首ldbestoredat-20oCuponreceiptwhiletheothers首ldbeat4oC.2.Foropenedkit:Whenthekitisopened,theremainingreagentsstillneedtobestoredaccordingtotheabovestoragecondition.Besides,pleasereturntheunusedwellstothefoilpouchcontainingthedesiccantpack,andresealalongentireedgeofzip-seal.Note:Itishighlyrecommendedtousetheremainingreagentswithin1monthprovidedthisiswithintheexpirationdateofthekit.[SAMPLECOLLECTIONANDSTORAGE]Serum-Allowsamplestoclotfortwohoursatroomtemperatureorovernightat4oCbeforecentrifugationfor20minutesatapproximately1000×g.Assayimmediatelyorstoresamplesinaliquotat-20oCor-80oC.Avoidrepeatedfreeze/thawcycles.Plasma-CollectplasmausingEDTAorheparinasananticoagulant.Centrifugesamplesfor15minutesat1000×gwithin30minutesofcollection.Removeplasmaandassayimmediatelyorstoresamplesinaliquotat-20oCor-80oC.Avoidrepeatedfreeze/thawcycles.Otherbiologicalfluids-Centrifugesamplesfor20minutesat1000×g.Removeparticulatesandassayimmediatelyorstoresamplesinaliquotat-20oCor-80oC.Avoidrepeatedfreeze/thawcycles.Note:1.Samplestobeusedwithin5daysmaybestoredat4oC,otherwisesamplesmustbestoredat-20oC（1month）or-80oC（2months）toavoidlossofbioactivityandcontamination.2.Samplehemolysiswillinfluencetheresult,sohemolyticspecimencannotbedetected.3.Whenperformingtheassay,bringsamplestoroomtemperature.[REAGENTPREPARATION]1.Bringallkitcomponentsandsamplestoroomtemperature（18-25oC）beforeuse.2.Standard-ReconstitutetheStandardwith1.0mLofStandardDiluent,keptfor10minutesatroomtemperature,shakegently（nottofoam）.Theconcentrationofthestandardinthestocksolutionis200ng/mL.Pleasefirstlydilutethestocksolutionto20ng/mLandthedilutedstandardservesasthehigheststandard（20ng/mL）.Thenprepare7tubescontaining0.5mLStandardDiluentandusethedilutedstandardtoproduceadoubledilutionseriesaccordingtothepictureshownbelow.Mixeachtubethoroughlybeforethenexttransfer.Setup7pointsofdilutedstandardsuchas20ng/mL,10ng/mL,5ng/mL,2.5ng/mL,1.25ng/mL,0.625ng/mL,0.312ng/mL,andthelastEPtubeswithStandardDiluentistheblankas0ng/mL.3.AssayDiluentAandAssayDiluentB-Dilute6mLofAssayDiluentAorBConcentrate（2×）with6mLofdeionizedordistilledwatertoprepare12mLofAssayDiluentAorB.（Infact,morethan6mLAssayDiluentAandAssayDiluentBarecontainedinthebottles.Therefore,ineverytest,pleasepreciselypipetterequiredamountofDiluentandmakedoubledilutioninanewcontainer.Thepreparedworkingdilutioncan'tbefrozen.）Tube123456789ng/mL200201052.51.250.6250.31204.DetectionReagentAandDetectionReagentB-BrieflyspinorcentrifugethestockDetectionAandDetectionBbeforeuse.DilutetotheworkingconcentrationwithworkingAssayDiluentAorB,respectively（1:100）.5.WashSolution-Dilute20mLofWashSolutionconcentrate（30×）with580mLofdeionizedordistilledwatertoprepare600mLofWashSolution（1×）.6.TMBsubstrate-Aspiratetheneededdosageofthesolutionwithsterilizedtipsanddonotdumptheresidualsolutionintothevialagain.Note:1.Makingserialdilutioninthewellsdirectlyisnotpermitted.2.Preparestandardwithin15minutesbeforeassay.Pleasedonotdissolvethereagentsat37oCdirectly.3.PleasecarefullyreconstituteStandardsorworkingDetectionReagentAandBaccordingtotheinstruction,andavoidfoamingandmixgentlyuntilthecrystalsarecompletelydissolved.Tominimizeimprecisioncausedbypipetting,usesmallvolumesandensurethatpipettorsarecalibrated.Itisrecommendedtosuckmorethan10μLforoncepipetting.4.ThereconstitutedStandards,DetectionReagentAandDetectionReagentBcanbeusedonlyonce.5.IfcrystalshaveformedintheWashSolutionconcentrate（30×）,warmtoroomtemperatureandmixgentlyuntilthecrystalsarecompletelydissolved.6.Contaminatedwaterorcontainerforreagentpreparationwillinfluencethedetectionresult.[SAMPLEPREPARATION]1.Uscn,Inc.isonlyresponsibleforthekititself,butnotforthesamplesconsumedduringtheassay.Theuser首ldcalculatethepossibleamountofthesamplesusedinthewholetest.Pleasereservesufficientsamplesinadvance.2.Pleasepredicttheconcentrationbeforeassaying.Ifvaluesforthesearenotwithintherangeofthestandardcurve,usersmustdeterminetheoptimalsampledilutionsfortheirparticularexperiments.3.Ifthesamplesarenotindicatedinthemanual,apreliminaryexperimenttodeterminethevalidityofthekitisnecessary.4.TissueorcellextractionsamplespreparedbychemicallysisbuffermaycauseunexpectedELISAresultsduetotheimpactsfromcertainchemicals.5.Duetothepossibilityofmismatchingbetweenantigenfromotheroriginandantibodyusedinourkits（e.g.,antibodytargetsconformationalepitoperatherthanlinearepitope）,somenativeorrecombinantproteinsfromothermanufacturersmaynotberecognizedbyourproducts.6.Influencedbythefactorsincludingcellviability,cellnumberorsamp領(lǐng)time,samplesfromcellculturesupernatantmaynotbedetectedbythekit.7.Freshsampleswithoutlongtimestorageisrecommendedforthetest.Otherwise,proteindegradationanddenaturalizationmayoccurinthosesamplesandfinallyleadtowrongresults.[ASSAYPROCEDURE]1.Determinewellsfordilutedstandard,blankandsample.Prepare7wellsforstandard,1wellforblank.Add100μLeachofdilutionsofstandard（readReagentPreparation）,blankandsamplesintotheappropriatewells.CoverwiththePlatesealer.Incubatefor2hoursat37oC.2.Removetheliquidofeachwell,don’twash.3.Add100μLofDetectionReagentAworkingsolutiontoeachwell.Incubatefor1hourat37oCaftercoveringitwiththePlatesealer.4.Aspiratethesolutionandwashwith350μLof1×WashSolutiontoeachwellusingasquirtbottle,multi-channelpipette,manifolddispenserorautowasher,andletitsitfor1~2minutes.Removetheremainingliquidfromallwellscompletelybysnappingtheplateontoabsorbentpaper.Totallywash3times.Afterthelastwash,removeanyremainingWashBufferbyaspiratingordecanting.Inverttheplateandblotitagainstabsorbentpaper.5.Add100μLofDetectionReagentBworkingsolutiontoeachwell.Incubatefor30minutesat37oCaftercoveringitwiththePlatesealer.6.Repeattheaspiration/washprocessfortotal5timesasconductedinstep4.7.Add90μLofSubstrateSolutiontoeachwell.CoverwithanewPlatesealer.Incubatefor15-25minutesat37oC（Don'texceed30minutes）.Protectfromlight.TheliquidwillturnbluebytheadditionofSubstrateSolution.8.Add50μLofStopSolutiontoeachwell.TheliquidwillturnyellowbytheadditionofStopsolution.Mixtheliquidbytappingthesideoftheplate.Ifcolorchangedoesnotappearuniform,gentlytaptheplatetoensurethoroughmixing.9.Removeanydropofwaterandfingerprintonthebottomoftheplateandconfirmthereisnobubbleonthesurfaceoftheliquid.Then,runthemicroplatereaderandconductmeasurementat450nmimmediately.Note:1.Assaypreparation:Keepappropriatenumbersofwellsfor1experimentandremoveextrawellsfrommicroplate.Restwells首ldberesealedandstoredat-20oC.2.SamplesorreagentsadditionPleaseusethefreshlypreparedStandard.Pleasecarefullyaddsamplestowellsandmixgentlytoavoidfoaming.Donottouchthewellwall.Foreachstepintheprocedure,totaldispensingtimeforadditionofreagentsorsamplestotheassayplate首ldnotexceed10minutes.Thiswillensureequalelapsedtimeforeachpipettingstep,withoutinterruption.Duplicationofallstandardsandspecimens,althoughnotrequired,isrecommended.Toavoidcross-contamination,changepipettetipsbetweenadditionsofstandards,samples,andreagents.Also,useseparatedreservoirsforeachreagent.3.Incubation:Toensureaccurateresults,properadhesionofplatesealersduringincubationstepsisnecessary.Donotallowwellstosituncoveredforextendedperiodsbetweenincubationsteps.Oncereagentsareaddedtothewellstrips,DONOTletthestripsDRYatanytimeduringtheassay.Incubationtimeandtemperaturemustbecontrolled.4.Washing:Thewashprocedureiscritical.Completeremovalofliquidateachstepisessentialforgoodperformance.Afterthelastwash,removeanyremainingWashSolutionbyaspiratingordecantingandremoveanydropofwaterandfingerprintonthebottomoftheplate.Insufficientwashingwillresultinpoorprecisionandfalseelevatedabsorbancereading.5.Control領(lǐng)ofreactiontime:ObservethechangeofcolorafteraddingTMBSubstrate（e.g.observationonceevery10minutes）,ifthecoloristoodeep,addStopSolutioninadvancetoavoidexcessivelystrongreactionwhichwillresultininaccurateabsorbancereading.6.TMBSubstrateiseasilycontaminated.Pleaseprotectitfromlight.7.Theenvironmenthumiditywhichislessthan60%mighthavesomeeffectsonthefinalperformance,therefore,ahumidifierisrecommendedtobeusedatthatcondition.[TESTPRINCIPLE]Themicrotiterplateprovidedinthiskithasbeenpre-coatedwithanantibodyspecifictouPAR.Standardsorsamplesarethenaddedtotheappropriatemicrotiterplatewellswithabiotin-conjugatedantibodypreparationspecificforuPAR.Next,AvidinconjugatedtoHorseradishPeroxidase（HRP）isaddedtoeachmicroplatewellandincubated.AfterTMBsubstratesolutionisadded,onlythosewellsthatcontainuPAR,biotin-conjugatedantibodyandenzyme-conjugatedAvidinwillexhibitachangeincolor.Theenzyme-substratereactionisterminatedbytheadditionofsulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm±10nm.TheconcentrationofuPARinthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.[CALCULATIONOFRESULTS]Averagetheduplicatereadingsforeachstandard,control,andsamplesandsubtracttheaveragezerostandardopticaldensity.Createastandardcurveonlog-loggraphpaper,withuPARconcentrationonthey-axisandabsorbanceonthex-axis.Drawthebestfitstraightlinethroughthestandardpointsanditcanbedeterminedbyregressionanalysis.Usingsomeplotsoftware,forinstance,curveexpert1.30,isalsorecommended.Ifsampleshavebeendiluted,theconcentrationreadfromthestandardcurvemustbemultipliedbythedilutionfactor.[TYPICALDATA]Inordertomakethecalculationeasier,weplottheO.D.valueofthestandard（X-axis）againsttheknownconcentrationofthestandard（Y-axis）,althoughconcentrationistheindependentvariableandO.D.valueisthedependentvariable.However,theO.D.valuesofthestandardcurvemayvaryaccordingtotheconditionsofassayperformance（e.g.operator,pipettingtechnique,washingtechniqueortemperatureeffects）,plottinglogofthedatatoestablishstandardcurveforeachtestisrecommended.Typicalstandardcurvebelowisprovidedforreferenceonly.TypicalStandardCurveforHumanuPARELISA.[DETECTIONRANGE]0.312-20ng/mL.ThestandardcurveconcentrationsusedfortheELISA’swere20ng/mL,10ng/mL,5ng/mL,2.5ng/mL,1.25ng/mL,0.625ng/mL,0.312ng/mL.[SENSITIVITY]TheminimumdetectabledoseofhumanuPARistypicallylessthan0.123ng/mL.Thesensitivityofthisassay,orLowerLimitofDetection（LLD）wasdefinedasthelowestproteinconcentrationthatcouldbedifferentiatedfromzero.Itwasdeterminedbyaddingtwostandarddeviationstothemeanopticaldensityvalueoftwentyzerostandardreplicatesandcalculatingthecorrespondingconcentration.[SPECIFICITY]ThisassayhashighsensitivityandexcellentspecificityfordetectionofhumanuPAR.Nosignificantcross-reactivityorinterferencebetweenhumanuPARandanalogueswasobserved.Note:Limitedbycurrentskillsandknowledge,itisimpossibleforustocompletethecross-reactivitydetectionbetweenhumanuPARandalltheanalogues,therefore,crossreactionmaystillexist.[RECOVERY]MatriceslistedbelowwerespikedwithcertainlevelofrecombinanthumanuPARandtherecoveryrateswerecalculatedbycomparingthemeasuredvaluetotheexpectedamountofuPARinsamples.MatrixRecoveryrange（%）Average（%）humanserum（n=5）91-10597humanEDTAplasma（n=5）95-10498humanheparinplasma（n=5）80-9688[LINEARITY]ThelinearityofthekitwasassayedbytestingsamplesspikedwithappropriateconcentrationofhumanuPARandtheirserialdilutions.Theresultsweredemonstratedbythepercentageofcalculatedconcentrationtotheexpected.Sample121418116humanserum（n=5）95-109%80-91%91-107%81-95%humanEDTAplasma（n=5）93-101%76-99%90-99%83-93%humanheparinplasma（n=5）78-96%94-107%101-106%89-98%[PRECISION]Intra-assayPrecision（Precisionwithinanassay）:3sampleswithlow,middleandhighlevelhumanuPARweretested20timesononeplate,respectively.Inter-assayPrecision（Precisionbetweenassays）:3sampleswithlow,middleandhighlevelhumanuPARweretestedon3differentplates,8replicatesineachplate.CV（%）=SD/meanX100Intra-Assay:CV<10%Inter-Assay:CV<12%[STABILITY]ThestabilityofELISAkitisdeterminedbythelossrateofactivity.Thelossrateofthiskitislessthan5%withintheexpirationdateunderappropriatestoragecondition.Thelossratewasdeterminedbyacceleratedthermaldegradationtest.Keepthekitat37oCfor3days,andcompareO.D.valuesofthekitkeptat37oCwiththatofatrecommendedtemperature.（referringfromChinaBiologicalProductsStandard,whichwascalculatedbytheArrheniusequation.ForELISAkit,1daystorageat37oCcanbeconsideredas2monthsat4oC,whichmeans3daysat37oCequa領(lǐng)6monthsat4oC）.Note:Tominimizeextrainfluenceontheperformance,operationproceduresandlabconditions,especiallyroomtemperature,airhumidity,incubatortemperature首ldbestrictlycontrolled.Itisalsostronglysuggestedthatthewholeassayisperformedbythesameoperatorfromthebeginningtotheend.[ASSAYPROCEDURESUMMARY]1.Prepareallreagents,samplesandstandards;2.Add100μLstandardorsampletoeachwell.Incubate2hoursat37oC;3.Add100μLpreparedDetectionReagentA.Incubate1hourat37oC;4.Aspirateandwash3times;5.Add100μLpreparedDetectionReagentB.Incubate30minutesat37oC;6.Aspirateandwash5times;7.Add90μLSubstrateSolution.Incubate15-25minutesat37oC;8.Add50μLStopSolution.Readat450nmimmediately.[IMPORTANTNOTE]1.Limitedbythecurrentconditionandscientifictechnology,wecan'tcompletelyconductthecomprehensiveidentificationandanalysisontherawmaterialprovidedbysuppliers.Sotheremightbesomequalitativeandtechnicalriskstousethekit.2.Thefinalexperimentalresultswillbecloselyrelatedtovalidityoftheproducts,operationskillsoftheendusersandtheexperimentalenvironments.Pleasemakesurethatsufficientsamplesareavailable.3.Kitsfromdifferentbatchesmaybealittledifferentindetectionrange,sensitivityandcolordevelopingtime.Pleaseperformtheexperimentexactlyaccordingtotheinstructionattachedinkitwhileelectroniconesfromourwebsite（www.uscnk.us;www.uscnk.cn;www.uscnk.com）isonlyforinformation.4.Donotmixorsubstitutereagentsfromonekitlottoanother.Useonlythereagentssuppliedbymanufacturer.5.Protectallreagentsfromstronglightduringstorageandincubation.Allthebottlecapsofreagents首ldbecoveredtightlytopreventtheevaporationandcontaminationofmicroorganism.6.Theremaybesomefoggysubstanceinthewellswhentheplateisopenedatthefirsttime.Itwillnothaveanyeffectonthefinalassayresults.Donotremovemicrotiterplatefromthestoragebaguntilneeded.7.Wrongoperationsduringthereagentspreparationandloading,aswellasincorrectparametersettingfortheplatereadermayleadtoincorrectresults.Amicroplateplatereaderwithabandwidthof10nmorlessandanopticaldensityrangeof0-3O.D.orgreaterat450±10nmwavelengthisacceptableforuseinabsorbancemeasurement.Pleasereadtheinstructioncarefullyandadjusttheinstrumentpriortotheexperiment.Formoreinformation,pleaserefertotheoperationvideo（http://www.uscnk.com/homepage/operate-elisa.htm）.8.Eventhesameoperatormightgetdifferentresultsintwoseparateexperiments.Inordertogetbetterreproducibleresults,theoperationofeverystepintheassay首ldbecontrolled.Furthermore,apreliminaryexperimentbeforeassayforeachbatchisrecommended.9.EachkithasbeenstrictlypassedQ.Ctest.However,resultsfromendusersmightbeinconsistentwithourin-housedataduetosomeunexpectedtransportationconditionsordifferentlabequipments.Intra-assayvarianceamongkitsfromdifferentbatchesmightarisefromabovefactors,too.10.Kitsfromdifferentmanufacturerswiththesameitemmightproducedifferentresults,sincewehaven’tcomparedourproductswithothermanufacturers.11.Validperiod:sixmonths.12.Theinstructionmanualalsosuitsforthekitof48T,butallreagentsof48Tkitarereducedbyhalf.[PRECAUTION]TheStopSolutionsuggestedforusewiththiskitisanacidsolution.Weareye,hand,face,andclothingprotectionwhenusingthismaterial.[詳細]

  2018-10-01 10:01

  產(chǎn)品樣冊

  |

  • 
  • 
  • 
  • 

• （原裝進口）人尿激酶型纖溶酶原激活物受體（uPAR）Elisa試劑盒說明書

  QuantikineHumanuPARImmunoassayCatalogNumberDUP00Forthequantitativedeterminationofhumanurokinase-typeplasminogenactivatorreceptor（uPAR）concentrationsincellculturesupernates,serum,plasma,andurine.Thispackageinsertmustbereadinitsentiretybeforeusingthisproduct.FORRESEARCHUSEONLY.NOTFORUSEINDIAGNOSTICPROCEDURES.TABLEOFCONTENTSContentsPageINTRODUCTION2PRINCIPLEOFTHEASSAY..................................3LIMITATIONSOFTHEPROCEDURE3MATERIALSPROVIDED....................................3STORAGE4OTHERSUPPLIESREQUIRED.................................4PRECAUTION4SAMPLECOLLECTIONANDSTORAGE............................5SAMPLEPREPARATION5REAGENTPREPARATION...................................6ASSAYPROCEDURE7ASSAYPROCEDURESUMMARY...............................8CALCULATIONOFRESULTS9TYPICALDATA.........................................9TECHNICALHINTS10PRECISION...........................................10LINEARITY11SENSITIVITY..........................................11CALIBRATION11SAMPLEVALUES.......................................12SPECIFICITY13REFERENCES.........................................14PLATELAYOUT15MANUFACTUREDANDDISTRIBUTEDBY:R&DSystems,Inc.TELEPHONE:（800）343-7475614McKinleyPlaceNE（612）379-2956Minneapolis,MN55413FAX:（612）656-4400UnitedStatesofAmericaE-MAIL:info@RnDSystems.comDISTRIBUTEDBY:R&DSystemsEurope,Ltd.19BartonLaneTELEPHONE:+44（0）1235529449AbingdonScienceParkFAX:+44（0）1235533420Abingdon,OX143NBE-MAIL:info@RnDSystems.co.ukUnitedKingdomR&DSystemsChinaCo.Ltd.24A1HuaMinEmpirePlazaTELEPHONE:+86（21）52380373726WestYanAnRoadFAX:+86（21）52371001ShanghaiPRC200050E-MAIL:info@RnDSystemsChina.com.cnINTRODUCTIONUrokinase-typeplasminogenactivatorreceptor（uPAR,CD87）isacellsurfacereceptorthatbindsurokinase-typeplasminogenactivator（uPA）withhighaffinity,therebyfacilitatingthepericellularactivationofplasminogen（seereferences1and2forreviews）.uPAR,uPAandplasminogenactivatorinhibitor-1（PAI-1）,formatriadwithmultiplefunctions,includingregulationofcellattachment,migration,proliferationanddifferentiation,bybothproteolyticandnonproteolyticmechanisms（2）.uPARisanchoredtoextracellularsurfacesthroughaglycosylphosphatidylinositol（GPI）linkage,withnotransmembranedomain（3）.uPARissynthesizedasa335-amino-acidpolypeptidethatincludesa22-residuesignalpeptide（4）.A30-residuepeptideiscleavedfromtheC-terminusduringadditionoftheGPIanchor（3）.WithlossofthesignalpeptideandtheC-terminalpeptide,thematureproteinhas283aminoacids.Itisvariablyglycosylated,increasingitsmassfromabout31kDafortheproteinbackbonetoasmuchas55kDaforthematureglycoprotein（5）.Pro-uPA,asingle-chainprotein,isactivatedtoadisulfide-linked,two-chainproteinbyproteolyticcleavagebyplasmin（1,2,6）.Eitherpro-uPAortheactivetwo-chainuPAbindwithhighaffinitytouPAR.Thus,tracesofplasminactivatepro-uPAtouPA,leadingtoincreasinggenerationofplasmininapositivefeedbackloopthatisamplifiedbyuPAR.Whiletheinitiatingeventisnotclear,theeffectisthegenerationofplasminatthecellsurface,whereitdegradestheextracellularmatrixbyactivatingmatrixmetalloproteinases.Thisappearstobeakeyeventintumorinvasivenessandmetastasisandinmigrationofcellsingeneral（1,2,7）.ThefunctionsoftheuPA/uPARsystemare,however,moreextensivethanmediationofplasminformation,andtheyincludenon-proteolyticfunctions（1,2）.uPA/uPARislocalizedtofocalcontactpointsofcellswithinthesubstratum.Thesesitesincludeintracellularvinculinandtheextracellularadhesionmoleculevitronectin,suggestingdirectadhesivefunctionsandintracellularsignal領(lǐng)functionsforuPAR.uPARbindstointegrins,anditcontainschemotacticactivityinitssingleprotease-sensitiveregion.uPARhasbeenmeasuredinhumanplasma（7-9）.SolubleuPARisgeneratedbyremovaloftheGPIanchorbyanendogenousphospholipaseD,freeinguPARofitssurfaceattachment（10）.uPARiselevatedinplasmaofpatientswithparoxysmalnocturnalhemoglobinuria（7,8）,amanifestationofaninabilitytoaddGPIanchorstoproteins.IthasbeenpostulatedthattherealsomaybesolubleuPARduetoalternativesplicingoftheprimarytranscript（1）,ashasbeendemonstratedformouseuPAR（11）.uPARhasbeenidentifiedinurine,wherethelevelisaconsistentfractionofcreatinineconcentration（12）.TheQuantikineHumanuPARImmunoassayisa4.5hoursolid-phaseELISAdesignedtomeasurehumanuPARincellculturesupernates,serum,plasma,andurine.ItcontainsNS0-expressedrecombinanthumanuPARandantibodiesraisedagainsttherecombinantfactor.Ithasbeenshowntoaccuratelyquantitatetherecombinantfactor.ResultsobtainedusingnaturalhumanuPARshowedlinearcurvesthatwereparalleltothestandardcurvesobtainedusingtheQuantikinekitstandards.TheseresultsindicatethatthiskitcanbeusedtodeterminerelativemassvaluesofnaturalhumanuPAR.2PRINCIPLEOFTHEASSAYThisassayemploysthequantitativesandwichenzymeimmunoassaytechnique.AmonoclonalantibodyspecificforuPARhasbeenpre-coatedontoamicroplate.StandardsandsamplesarepipettedintothewellsandanyuPARpresentisboundbytheimmobilizedantibody.Afterwashingawayanyunboundsubstances,anenzyme-linkedpolyclonalantibodyspecificforuPARisaddedtothewells.Followingawashtoremoveanyunboundantibody-enzymereagent,asubstratesolutionisaddedtothewellsandcolordevelopsinproportiontotheamountofuPARboundintheinitialstep.Thecolordevelopmentisstoppedandtheintensityofthecolorismeasured.LIMITATIONSOFTHEPROCEDUREFORRESEARCHUSEONLY.NOTFORUSEINDIAGNOSTICPROCEDURES.Thekit首ldnotbeusedbeyondtheexpirationdateonthekitlabel.Donotmixorsubstitutereagentswithotherlotsorsources.Ifsamplesfalloutsidethedynamicrangeoftheassay,furtherdilutethesampleswithCalibratorDiluentandrepeattheassay.Anyvariationinstandarddiluent,operator,pipettingtechnique,washingtechnique,incubationtimeortemperature,andkitagecancausevariationinbinding.Thisassayisdesignedtoeliminateinterferencebyligandsandotherproteinspresentinbiologicalsamples.UntilallfactorshavebeentestedintheQuantikineImmunoassay,thepossibilityofinterferencecannotbeexcluded.MATERIALSPROVIDEDuPARMicroplate（Part890714）-96wellpolystyrenemicroplate（12stripsof8wells）coatedwithamousemonoclonalantibodyagainstuPAR.uPARConjugate（Part890715）-21mLofpolyclonalantibodyagainstuPARconjugatedtohorseradishperoxidasewithpreservatives.uPARStandard（Part890716）-40ngofrecombinanthumanuPARinabufferedproteinbasewithpreservatives;lyophilized.AssayDiluentRD1W（Part895117）-11mLofabufferedproteinbasewithpreservatives.CalibratorDiluentRD6P（Part895118）-21mLofanimalserumwithpreservatives.WashBufferConcentrate（Part895003）-21mLofa25-foldconcentratedsolutionofbufferedsurfactantwithpreservatives.ColorReagentA（Part895000）-12.5mLofstabilizedhydrogenperoxide.ColorReagentB（Part895001）-12.5mLofstabilizedchromogen（tetramethylbenzidine）.StopSolution（Part895032）-6mLof2Nsulfuricacid.PlateCovers-4adhesivestrips.3STORAGEUnopenedKitStoreat2-8°C.Donotusepastkitexpirationdate.Opened/ReconstitutedReagentsDilutedWashBufferMaybestoredforupto1monthat2-8°C.*StopSolutionAssayDiluentRD1WCalibratorDiluentRD6PConjugateUnmixedColorReagentAUnmixedColorReagentBStandardMicroplateWellsReturnunusedwellstothefoilpouchcontainingthedesiccantpack,resealalongentireedgeofzip-seal.Maybestoredforupto1monthat2-8°C.**Providedthisiswithintheexpirationdateofthekit.OTHERSUPPLIESREQUIREDMicroplatereadercapableofmeasuringabsorbanceat450nm,withthecorrectionwavelengthsetat540nmor570nm.Pipettesandpipettetips.Deionizedordistilledwater.Squirtbottle,manifolddispenser,orautomatedmicroplatewasher.500mLgraduatedcylinder.Testtubesfordilution.HumanuPARControls（optional;availablefromR&DSystems）.PRECAUTIONTheStopSolutionprovidedwiththiskitisanacidsolution.Weareye,hand,face,andclothingprotectionwhenusingthismaterial.4SAMPLECOLLECTIONANDSTORAGECellCultureSupernates-Removeparticulatesbycentrifugationandassayimmediatelyoraliquotandstoresamplesat-20°C.Avoidrepeatedfreeze-thawcycles.Serum-Useaserumseparatortube（SST）andallowsamplestoclotfor30minutesbeforecentrifugationfor10minutesat1000xg.Removeserumandassayimmediatelyoraliquotandstoresamplesat-20°C.Avoidrepeatedfreeze-thawcycles.Plasma-CollectplasmausingheparinorEDTAasananticoagulant.Centrifugeat1000xgwithin30minutesofcollection.Assayimmediatelyoraliquotandstoresamplesat-20°C.Avoidrepeatedfreeze-thawcycles.Note:Citrateplasmahasnotbeenvalidatedforuseinthisassay.LipemicsamplesandsamplescontaininghighlevelsofhumanserumalbuminarenotsuitableforthemeasurementofhumanuPARwiththisassay.Urine-Asepticallycollectthefirsturineoftheday（mid-stream）,voideddirectlyintoasterilecontainer.Centrifugetoremoveparticulatematter.Assayimmediatelyoraliquotandstoreat-20°C.Avoidrepeatedfreeze-thawcycles.Formoreinformationoncollectingandhand領(lǐng)urinespecimens,refertotheNationalCommitteeforClinicalLaboratoryStandards（NCCLS）publicationGP16-T.SAMPLEPREPARATIONCellculturesupernates,serum,plasma,andurinesamplesrequireatleasta5-folddilution.Asuggested5-folddilutionis25Lofsample+100LofCalibratorDiluentRD6P.5REAGENTPREPARATIONBringallreagentstoroomtemperaturebeforeuse.WashBuffer-Ifcrystalshaveformedintheconcentrate,warmtoroomtemperatureandmixgentlyuntilthecrystalshavecompletelydissolved.Dilute20mLofWashBufferConcentrateintodeionizedordistilledwatertoprepare500mLofWashBuffer.SubstrateSolution-ColorReagentsAandB首ldbemixedtogetherinequalvolumeswithin15minutesofuse.Protectfromlight.200Loftheresultantmixtureisrequiredperwell.uPARStandard-ReconstitutetheuPARStandardwith1.0mLofdeionizedordistilledwater.Thisreconstitutionproducesastocksolutionof40,000pg/mL.Allowthestandardtositforaminimumof15minuteswithgentleagitationpriortomakingdilutions.Pipette450LofCalibratorDiluentRD6Pintothe4000pg/mLtube.Pipette200LofCalibratorDiluentRD6Pintotheremainingtubes.Usethestocksolutiontoproduceadilutionseries（below）.Mixeachtubethoroughlybeforethenexttransfer.The4000pg/mLstandardservesasthehighstandard.CalibratorDiluentRD6Pservesasthezerostandard（0pg/mL）.6ASSAYPROCEDUREBringallreagentsandsamplestoroomtemperaturebeforeuse.Itisrecommendedthatallsamples,standards,andcontrolsbeassayedinduplicate.1.Prepareallreagents,workingstandards,andsamplesasdirectedintheprevioussections.2.Removeexcessmicroplatestripsfromtheplateframe,returnthemtothefoilpouchcontainingthedesiccantpack,andreseal.3.Add100LofAssayDiluentRD1Wtoeachwell.4.Add50LofStandard,control,orsample*perwell.Coverwiththeadhesivestripprovided.Incubatefor2hoursatroomtemperature.Aplatelayoutisprovidedtorecordstandardsandsamplesassayed.5.Aspirateeachwellandwash,repeatingtheprocessthreetimesforatotaloffourwashes.Washbyfil領(lǐng)eachwellwithWashBuffer（400L）usingasquirtbottle,manifolddispenser,orautowasher.Completeremovalofliquidateachstepisessentialtogoodperformance.Afterthelastwash,removeanyremainingWashBufferbyaspiratingordecanting.Inverttheplateandblotitagainstcleanpapertowels.6.Add200LofuPARConjugatetoeachwell.Coverwithanewadhesivestrip.Incubatefor2hoursatroomtemperature.7.Repeattheaspiration/washasinstep5.8.Add200LofSubstrateSolutiontoeachwell.Incubatefor30minutesatroomtemperature.Protectfromlight.9.Add50LofStopSolutiontoeachwell.Thecolorinthewells首ldchangefrombluetoyellow.Ifthecolorinthewellsisgreenorifthecolorchangedoesnotappearuniform,gentlytaptheplatetoensurethoroughmixing.10.Determinetheopticaldensityofeachwellwithin30minutes,usingamicroplatereadersetto450nm.Ifwavelengthcorrectionisavailable,setto540nmor570nm.Ifwavelengthcorrectionisnotavailable,subtractreadingsat540nmor570nmfromthereadingsat450nm.Thissubtractionwillcorrectforopticalimperfectionsintheplate.Readingsmadedirectlyat450nmwithoutcorrectionmaybehigherandlessaccurate.*Samplesrequiredilution.SeeSamplePreparationsection.7ASSAYPROCEDURESUMMARY8CALCULATIONOFRESULTSAveragetheduplicatereadingsforeachstandard,control,andsampleandsubtracttheaveragezerostandardopticaldensity.Createastandardcurvebyreducingthedatausingcomputersoftwarecapableofgeneratingalog/logcurve-fit.Asanalternative,constructastandardcurvebyplottingthemeanabsorbanceforeachstandardonthey-axisagainsttheconcentrationonthex-axisanddrawabestfitcurvethroughthepointsonalog/loggraph.ThedatamaybelinearizedbyplottingthelogoftheuPARconcentrationsversusthelogoftheO.D.onalinearscale,andthebestfitlinecanbedeterminedbyregressionanalysis.Ifthesampleshavebeendiluted,theconcentrationreadfromthestandardcurvemustbemultipliedbythedilutionfactor.TYPICALDATAThisstandardcurveisprovidedfordemonstrationonly.Astandardcurve首ldbegeneratedforeachsetofsamplesassayed.9pg/mL062.5125250500100020004000O.D.0.0460.0440.0720.0730.1050.1050.1700.1710.3000.3040.5650.5451.0661.0752.1222.076Average0.0450.0720.1050.1700.3020.5551.0702.099Corrected___0.0270.0600.1250.2570.5101.0252.054TECHNICALHINTSWhenmixingorreconstitutingproteinsolutions,alwaysavoidfoaming.Toavoidcross-contamination,changepipettetipsbetweenadditionsofeachstandardlevel,betweensampleadditions,andbetweenreagentadditions.Also,useseparatereservoirsforeachreagent.Whenusinganautomatedplatewasher,addinga30secondsoakperiodfollowingtheadditionofwashbuffer,and/orrotatingtheplate180degreesbetweenwashstepsmayimproveassayprecision.Toensureaccurateresults,properadhesionofplatesealersduringincubationstepsisnecessary.SubstrateSolution首ldremaincolorlessuntiladdedtotheplate.KeeptheSubstrateSolutionprotectedfromlight.SubstrateSolution首ldchangefromcolorlesstogradationsofblue.StopSolution首ldbeaddedtotheplateinthesameorderastheSubstrateSolution.ThecolordevelopedinthewellswillturnfrombluetoyellowuponadditionoftheStopSolution.WellsthataregreenincolorindicatethattheStopSolutionhasnotmixedthoroughlywiththeSubstrateSolution.PRECISIONIntra-assayPrecision（Precisionwithinanassay）Threesamplesofknownconcentrationweretestedtwentytimesononeplatetoassessintra-assayprecision.Inter-assayPrecision（Precisionbetweenassays）Threesamplesofknownconcentrationweretestedinfortyseparateassaystoassessinter-assayprecision.Intra-assayPrecisionInter-assayPrecisionSample123123n202020404040Mean（pg/mL）8361593241279615462300Standarddeviation17.365.418144.378.9136CV（%）2.14.17.55.65.15.910LINEARITYToassessthelinearityoftheassay,samplesspikedwithhighconcentrationsofuPARweredilutedwithCalibratorDiluentRD6Ptoproducesampleswithvalueswithinthedynamicrangeoftheassay.Cellculturemedia*（n=4）Serum*（n=5）Heparinplasma*（n=5）EDTAplasma*（n=5）Urine*（n=5）1:2Average%ofExpected10396-112104101-105104101-10610194-105102Range（%）99-1051:4Average%ofExpected109101-115106103-111107104-110105101-108104Range（%）95-1121:8Average%ofExpected109101-111106103-112106102-110106103-113108Range（%）104-1131:16Average%ofExpected10999-11410599-11110598-11010498-111108Range（%）103-112*Sampleswerediluted5-foldpriortoassayasdirectedinSamplePreparation.SENSITIVITYTheminimumdetectabledose（MDD）ofuPARistypicallylessthan33pg/mL.TheMDDwasdeterminedbyaddingtwostandarddeviationstothemeanopticaldensityvalueoftwentyzerostandardreplicatesandcalculatingthecorrespondingconcentration.CALIBRATIONThisimmunoassayiscalibratedagainstahighlypurifiedNS0-expressedrecombinanthumanuPARproducedatR&DSystems.11SAMPLEVALUESSerum/Plasma/Urine-SamplesfromapparentlyhealthyvolunteerswereevaluatedforthepresenceofuPARinthisassay.Nomedicalhistorieswereavailableforthedonorsusedinthisstudy.SampleTypeMean（pg/mL）Range（pg/mL）Serum（n=60）23701195-4415Heparinplasma（n=35）2165977-5347EDTAplasma（n=35）1871864-3829Urine（n=35）2975691-6098CellCultureSupernates-Humanperipheralbloodmononuclearcells（5x106cells/mL）wereculturedinRPMIsupplementedwith5%fetalcalfserum,50M-mercaptoethanol,2mML-glutamine,100U/mLpenicillin,and100g/mLstreptomycinsulfate.Thecellswereculturedunstimulatedorstimulatedwith10g/mLPHA.AliquotsofthecellculturesupernateswereremovedandassayedforlevelsofnaturaluPAR.ConditionDay1（pg/mL）Day5（pg/mL）Unstimulated12282123Stimulated10,005689512SPECIFICITYThisassayrecognizesrecombinantandnaturalhumanuPAR.Thefactorslistedbelowwerepreparedat50ng/mLinCalibratorDiluentRD6Pandassayedforcross-reactivity.Preparationsofthefollowingfactorsat50ng/mLinamid-rangerecombinanthumanuPARcontrolwereassayedforinterference.Nosignificantcross-reactivityorinterferencewasobserved.RecombinanthumanuPAdoesnotcross-reactinthisassaybutdoesinterfereatconcentrationsgreaterthan3000pg/mL.13Recombinanthuman:ANGARCNTF-ECGFEGFEpoFGFacidicFGFbasicFGF-4FGF-5FGF-6Flt-3LigandG-CSFGM-CSFsgp130GROGROGROHB-EGFHGFIFN-IGF-IIGF-IIIL-1IL-1IL-1raIL-1sRIIL-1sRIIIL-2IL-2sRIL-3IL-3sRIL-4IL-4sRIL-5IL-5sRIL-5sRIL-6IL-6sRIL-7IL-8IL-9IL-10IL-11IL-12IL-13KGF（FGF-7）LAP（TGF-1）LIFM-CSFMCP-1MIP-1MIP-1-NGFOSMPD-ECGFPDGF-AAPDGF-ABPDGF-BBPTNRANTESSCFSLPITGF-TGF-1TGF-2TGF-3TGF-sRIITNF-TNF-sTNFRIsTNFRIIVEGFRecombinantmouse:GM-CSFIL-1IL-1IL-3IL-4IL-5IL-6IL-7IL-9IL-10IL-13LIFMIP-1MIP-1SCFTNF-uPARRecombinantamphibian:TGF-5Naturalproteins:bovineFGFacidicbovineFGFbasichumanPDGFporcinePDGFhumanTGF-1porcineTGF-1REFERENCES1.Dear,A.E.andR.L.Medcalf（1998）Eur.J.Biochem.252:185.2.Blasi,F.（1997）ImmunlogyToday18:415.3.Ploug,M.etal.（1991）J.Biol.Chem.266:1926.4.Roldan,A.L.（1990）EMBOJ.9:467.5.Behrendt,N.（1990）J.Biol.Chem.265:6453.6.Blasi,F.etal.（1987）J.CellBiol.104:801.7.Ronne,E.etal.（1995）Br.J.Haematol.89:576.8.Ploug,M.etal.（1992）Blood79:1447.9.Stephens,R.W.etal.（1999）J.Natl.CancerInst.91:869.10.Wilhelm,O.G.etal.（1999）J.CellularPhysiol.180:225.11.Kristensen,P.etal.（1991）J.CellBiol.115:1763.12.Sier,C.F.M.etal.（1999）Lab.Invest.79:717.14PLATELAYOUTUsethisplatelayouttorecordstandardsandsamplesassayed.2010R&DSystems,Inc.12.99750440.57/10[詳細]

  2018-10-01 10:01

  產(chǎn)品樣冊

  |

  • 
  • 
  • 
  • 

• 大鼠尿激酶型纖溶酶原激活物(uPA)ELISA試劑盒

  大鼠尿激酶型纖溶酶原激活物(uPA)ELISA試劑盒[詳細]

  2013-12-11 00:00

  應(yīng)用文章

  |

  • 
  • 
  • 
  • 

• 人可溶性尿激酶型纖溶酶原激活物受體試劑盒使用方法

  人可溶性尿激酶型纖溶酶原激活物受體試劑盒使用方法[詳細]

  2016-09-06 00:00

  產(chǎn)品樣冊

  |

  • 
  • 
  • 
  • 

• 人可溶性尿激酶型纖溶酶原激活物受體 試劑盒使用方法

  本試劑盒僅供研究使用。檢測范圍：96T[詳細]

  2018-10-01 10:00

  產(chǎn)品樣冊

  |

  • 
  • 
  • 
  • 

• 小鼠尿激酶型纖溶酶原激活物受體(PLAUR/uPAR)ELISA試劑盒elisa試劑盒說明書

  小鼠尿激酶型纖溶酶原激活物受體(PLAUR/uPAR)ELISA試劑盒elisa試劑盒說明書[詳細]

  2024-10-08 05:27

  期刊論文

  |

  • 
  • 
  • 
  • 

• 小鼠尿激酶型纖溶酶原激活物受體（uPA-R）ELISA試劑盒

  試驗原理：uPA-R試劑盒是固相夾心法酶聯(lián)免疫吸附實驗（ELISA）.已知uPA-R濃度的標準品、未知濃度的樣品加入微孔酶標板內(nèi)進行檢測。先將uPA-R和生物素標記的抗體同時溫育。洗滌后，加入親和素標記過的HRP。再經(jīng)過溫育和洗滌，去除未結(jié)合的酶結(jié)合物，然后加入底物A、B，和酶結(jié)合物同時作用。產(chǎn)生顏色。顏色的深淺和樣品中uPA-R的濃度呈比例關(guān)系。試劑盒內(nèi)容及其配制試劑盒成份（2-8℃保存）96孔配置48孔配置配制96/48人份酶標板1塊板（96T）半塊板（48T）即用型塑料膜板蓋1塊半塊即用型標準品：40ng/ml1瓶（0.6ml）1瓶（0.3ml）按說明書進行稀稀空白對照1瓶（1.0ml）1瓶（0.5ml）即用型標準品稀釋緩沖液1瓶（4.0ml）1瓶（2.0ml）即用型生物素標記的抗uPA-R抗體1瓶（6.0ml）1瓶（3.0ml）即用型親和鏈酶素-HRP1瓶（8.0ml）1瓶（4.0ml）即用型洗滌緩沖液1瓶（20ml）1瓶（10ml）按說明書進行稀釋底物A1瓶（6.0ml）1瓶（3.0ml）即用型底物B1瓶（6.0ml）1瓶（3.0ml）即用型終止液1瓶（6.0ml）1瓶（3.0ml）即用型標本稀釋液1瓶（12ml）1瓶（6.0ml）即用型自備材料1．蒸餾水。2．加樣器：5ul、10ul、50ul、100ul、200ul、500ul、1000ul。3．振蕩器及磁力攪拌器等。小鼠尿激酶型纖溶酶原激活物受體(uPA-R)ELISA試劑盒安全性1．避免直接接觸終止液和底物A、B。一旦接觸到這些液體，請盡快用水沖洗。2．實驗中不要吃喝、抽煙或使用化妝品。3．不要用嘴吸取試劑盒里的任何成份。操作注意事項1．試劑應(yīng)按標簽說明書儲存，使用前恢復(fù)到室溫。稀稀過后的標準品應(yīng)丟棄，不可保存。2．實驗中不用的板條應(yīng)立即放回包裝袋中，密封保存，以免變質(zhì)。3．不用的其它試劑應(yīng)包裝好或蓋好。不同批號的試劑不要混用。保質(zhì)前使用。4．使用一次性的吸頭以免交叉污染，吸取終止液和底物A、B液時，避免使用帶金屬部分的加樣器。5．使用干凈的塑料容器配置洗滌液。使用前充分混勻試劑盒里的各種成份及樣品。6．洗滌酶標板時應(yīng)充分拍干，不要將吸水紙直接放入酶標反應(yīng)孔中吸水。7．底物A應(yīng)揮發(fā)，避免長時間打開蓋子。底物B對光敏感，避免長時間暴露于光下。避免用手接觸，有毒。實驗完成后應(yīng)立即讀取OD值。8．加入試劑的順序應(yīng)一致，以保證所有反應(yīng)板孔溫育的時間一樣。9．按照說明書中標明的時間、加液的量及順序進行溫育操作。樣品收集、處理及保存方法1、血清-----操作過程中避免任何細胞刺激。使用不含熱原和內(nèi)毒素的試管。收集血液后，1000×g離心10分鐘將血清和紅細胞迅速小心地分離。2、血漿-----EDTA、檸檬酸鹽、肝素血漿可用于檢測。1000×g離心30分鐘去除顆粒。3、細胞上清液---1000×g離心10分鐘去除顆粒和聚合物。4、組織勻漿-----將組織加入適量生理鹽水搗碎。1000×g離心10分鐘，取上清液5、保存------如果樣品不立即使用，應(yīng)將其分成小部分-70℃保存，避免反復(fù)冷凍。盡可能的不要使用溶血或高血脂血。如果血清中大量顆粒，檢測前先離心或過濾。不要在37℃或更高的溫度加熱解凍。應(yīng)在室溫下解凍并確保樣品均勻地充分解凍。小鼠尿激酶型纖溶酶原激活物受體(uPA-R)ELISA試劑盒試劑的準備1．標準品：標準品的系列稀釋應(yīng)在實驗時準備，不能儲存。稀釋前將標準品振蕩混勻。稀釋比例按下表中進行：40ng/ml（6號標準品）原倍濃度不用稀釋直接加入50ul。20ng/ml（5號標準品）100ul的原倍標準品加入100ul的標準品稀釋液10ng/ml（4號標準品）100ul的5號標準品加入100ul的標準品稀釋液5.0ng/ml（3號標準品）100ul的4號標準品加入100ul的標準品稀釋液2.5ng/ml（2號標準品）100ul的3號標準品加入100ul的標準品稀釋液1.25ng/ml（1號標準品）100ul的2號標準品加入100ul的標準品稀釋液0ng/ml（空白對照）原始濃度不用稀釋直接加入50ul。2．洗滌緩沖液（50×）的稀釋：蒸餾水50倍稀釋。操作步驟1．使用前，將所有試劑充分混勻。不要使液體產(chǎn)生大量的泡沫，以免加樣時加入大量的氣泡，產(chǎn)生加樣上的誤差。2．根據(jù)待測樣品數(shù)量加上標準品的數(shù)量決定所需的板條數(shù)。每個標準品和空白孔建議做復(fù)孔。每個樣品根據(jù)自己的數(shù)量來定，能使用復(fù)孔的盡量做復(fù)孔。標本用標本稀釋液1：1稀釋后加入50ul于反應(yīng)孔內(nèi)。3．加入稀釋好后的標準品50ul于反應(yīng)孔、加入待測樣品50ul于反應(yīng)孔內(nèi)。立即加入50ul的生物素標記的抗體。蓋上膜板，輕輕振蕩混勻，37℃溫育1小時。4．甩去孔內(nèi)液體，每孔加滿洗滌液，振蕩30秒，甩去洗滌液，用吸水紙拍干。重復(fù)此操作3次。如果用洗板機洗滌，洗滌次數(shù)增加一次。5．每孔加入60ul的親和鏈酶素-HRP，輕輕振蕩混勻，37℃溫育30分鐘。6．甩去孔內(nèi)液體，每孔加滿洗滌液，振蕩30秒，甩去洗滌液，用吸水紙拍干。重復(fù)此操作3次。如果用洗板機洗滌，洗滌次數(shù)增加一次。7．每孔加入底物A、B各50ul，輕輕振蕩混勻，37℃溫育10分鐘。避免光照。8．取出酶標板，迅速加入50ul終止液，加入終止液后應(yīng)立即測定結(jié)果。9．在450nm波長處測定各孔的OD值。建議使用的實驗方案標準品濃度（ng/ml）A4040樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品B2020樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品C1010樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品D5.05.0樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品E2.52.5樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品F1.251.25樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品G00樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品H樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品局限6號標準品以上的結(jié)果為非線性的，根據(jù)此標準曲線無法得到極ng確的結(jié)果。試劑盒性能1.靈敏度：Z小的檢測濃度小于1號標準品。稀釋度的線性。樣品線性回歸與預(yù)期濃度相關(guān)系數(shù)R值為0.990。2.特異性：不與其它細胞因子反應(yīng)。3.重復(fù)性：板內(nèi)、板間變異系數(shù)均小于10%。結(jié)果判斷與分析1、儀器值：于波長450nm的酶標儀上讀取各孔的OD值2、以吸光度OD值為縱坐標（Y），相應(yīng)的uPA-R標準品濃度為橫坐標（X），做得相應(yīng)的曲線，樣品的uPA-R含量可根據(jù)其OD值由標準曲線換算出相應(yīng)的濃度。3、檢測值范圍：0-40ng/ml4、敏感度：0.1ng/ml[詳細]

  2018-09-27 10:00

  產(chǎn)品樣冊

  |

  • 
  • 
  • 
  • 

• 人可溶性尿激酶型纖溶酶原激活物受體酶聯(lián)免疫分析試劑盒使用方法

  本試劑盒僅供研究使用。檢測范圍：96T[詳細]

  2018-12-30 10:00

  產(chǎn)品樣冊

  |

  • 
  • 
  • 
  • 

• 體液尿激酶（UROKINASE）活性熒光定量檢測試劑盒說明書

  主要用途是一種旨在通過三肽化合物底物p-ERG-pNA受到尿激酶的水解，釋放黃色對硝基苯胺，產(chǎn)生吸光峰值的變化，即采用比色法來測定細胞裂解萃取樣品中酶活性的權(quán)威而經(jīng)典的技術(shù)方法。該技術(shù)由大師級科學(xué)家精心研制、成功實驗證明的。其適用于各種細胞裂解萃取液樣品（動物、人體等）尿激酶的活性檢測。產(chǎn)品嚴格無菌，即到即用，操作簡捷，性能穩(wěn)定。技術(shù)背景尿激酶（urokinase；UK；EC3.4.21.73），又稱為尿激酶型纖維蛋白溶酶原激活劑（urokinase-typePlasminogenActivator；uPA），屬于絲氨酸蛋白酶，存在于血液、細胞外基質(zhì)和尿液中。尿激酶由411氨基酸構(gòu)成，分子量為54KD，有3個結(jié)構(gòu)域：絲氨酸蛋白酶結(jié)構(gòu)域、kringle結(jié)構(gòu)域和生長因子結(jié)構(gòu)域。其主要的生理性底物為纖維蛋白溶酶原（Plasminogen），即纖維蛋白溶酶（plasmin）的酶原形式（zymogen），切離部位為Cys-Pro-Gly-Arg⊥Val-Val-Gly-Gly-Cys。一旦切離后激活，纖維蛋白溶酶參與細胞蛋白水解過程，包括血栓溶解（thrombolysis）和細胞外基質(zhì)降解等。尿激酶與組織重構(gòu)、修復(fù)、血管形成（angiogenesis）性疾病和腫瘤擴散有關(guān)。尿激酶的YZ劑是絲氨酸蛋白酶YZ劑（serpins），即纖維蛋白溶酶原激活劑YZ劑（PlasminogenActivatorInhibitor；PAI）?；谌斯ず铣傻娜幕衔锏孜飌-EGR-pNA（pyro-Glu-Gly-Arg-p-Nithoaniline；焦谷氨酸酰甘氨酰精氨酰對硝基苯胺）受到尿激酶的水解，釋放有色對硝基苯胺，在分光光度儀（405nm波長）下，測定吸光峰值的變化，來定量測定尿激酶的活性。尿激酶反應(yīng)系統(tǒng)為：產(chǎn)品內(nèi)容清理液（ReagentA）毫升裂解液（ReagentB）毫升緩沖液（ReagentC）毫升陰性液（ReagentD）毫升底物液（ReagentE）微升產(chǎn)品說明書1份保存方式保存清理液（ReagentA）在4℃冰箱里；其余的保存在-20℃冰箱里；底物液（ReagentE）避免光照和反復(fù)凍融；有效保證6月用戶自備15[詳細]

  2018-11-08 10:00

  產(chǎn)品樣冊

  |

  • 
  • 
  • 
  • 

• 人骨粘連蛋白(ON)ELISA試劑盒

  人骨粘連蛋白(ON)ELISA試劑盒[詳細]

  2013-12-06 00:00

  選購指南

  |

  • 
  • 
  • 
  • 

• 人一氧化氮（NO）ELISA試劑盒

  電話：021-6533363955229872網(wǎng)址：http://www.westang.com人一氧化氮（NO）ELISA試劑盒（用于血清、血漿、細胞培養(yǎng)上清液和其它生物體液內(nèi)）原理本實驗采用雙抗體夾心ABC-ELISA法。用抗人NO單抗包被于酶標板上，標準品和樣品中的NO與單抗結(jié)合，加入生物素化的抗人NO，形成免疫復(fù)合物連接在板上，辣根過氧化物酶標記的Streptavidin與生物素結(jié)合，加入底物工作液顯藍色，Z后加終止液硫酸，在450nm處測OD值，NO濃度與OD值成正比，可通過繪制標準曲線求出標本中NO濃度。試劑盒組成（2-8℃保存）酶標板（CoatedWells）96孔酶標抗體工作液（EnzymeConjugate）12ml10×標本稀釋液（SampleBuffer）12ml20×濃縮洗滌液（WashBuffer）50ml標準品（Standards）：1000μM/瓶2瓶底物工作液（TMBSolution）12ml**抗體工作液（BiotinylatedAntibody）12ml終止液（StopSolution）12ml準備試劑與收集血樣1.收集標本：血清、血漿（EDTA）、細胞培養(yǎng)上清液、組織勻漿等盡早檢測，2-8℃保存48小時；更長時間須冷凍（-20℃或-70℃）保存，避免反復(fù)凍融。2.標準品液配制：使用前加入1ml蒸餾水混勻，配成1000μM的溶液。設(shè)標準管8管，**管加標本稀釋液900ul，第二至第八管加入標本稀釋液500ul。在**管中加入1000μM的標準品溶液100ul混勻后用加樣器吸出500ul，移至第二管。如此反復(fù)作對倍稀釋，從第七管中吸出500ul棄去。第八管為空白對照。3.10×標本稀釋液用蒸餾水作1:10倍稀釋（示例：1ml濃稀釋液+9ml蒸餾水）。4.洗滌液：用重蒸水1:20稀釋（示例：1ml濃縮洗滌液加入19ml的重蒸水）檢測程序1.加樣：每孔各加入標準品或待測樣品100ul，將反應(yīng)板充分混勻后置37℃120分鐘。2.洗板：用洗滌液將反應(yīng)板充分洗滌4-6次，向濾紙上印干。3.每孔中加入**抗體工作液100ul。將反應(yīng)板充分混勻后置37℃60分鐘。4.洗板：同前。5.每孔加酶標抗體工作液100ul。將反應(yīng)板置37℃30分鐘。6.洗板：同前。7.每孔加入底物工作液100ul，置37℃暗處反應(yīng)15分鐘。8.每孔加入100ul終止液混勻。9.30分鐘內(nèi)用酶標儀在450nm處測吸光值。結(jié)果計算與判斷1.所有OD值都應(yīng)減除空白值后再行計算。2.以標準品100、50、25、12.5、6.25、3.125、1.56、0μM為橫坐標，OD值為縱坐標，在坐標紙上作圖，畫出標準曲線。3.根據(jù)樣品OD值在該曲線圖上查出相應(yīng)NO含量。試劑盒性能1.靈敏度：Z小的NO檢測濃度小于1μM。2.特異性：可同時檢測重組或天然的人NO。不與人其它細胞因子有交叉反應(yīng)。3.重復(fù)性：板內(nèi)、板見變異系數(shù)均小于10％。注意事項1.以上標準孔及待測樣品均建議做復(fù)孔，每次測定應(yīng)同時做標準曲線。2.洗滌過程很關(guān)鍵。洗滌不充分將導(dǎo)致極ng確度誤差及OD值錯誤地升高。3.板條開封后剩余板條要再封好，保持板條干燥。4.本試劑盒宜置4oC冰箱保存。5.本試劑盒僅用于科研，不能用于臨床診斷！[詳細]

  2018-09-13 10:01

  產(chǎn)品樣冊

  |

  • 
  • 
  • 
  • 

• 人c-fos ELISA試劑盒

  人c-fosELISA試劑盒操作步驟僅供研究使用檢測范圍：96T3μg/L-120μg/L使用目的：人c-fosELISA試劑盒用于測定大鼠血清、血漿及相關(guān)液體樣本中內(nèi)皮素（ET）含量。實驗原理本試劑盒應(yīng)用雙抗體夾心法測定標本中大鼠內(nèi)皮素（ET）水平。用純化的大鼠內(nèi)皮素（ET）抗體包被微孔板，制成固相抗體，往包被單抗的微孔中依次加入內(nèi)皮素（ET），再與HRP標記的內(nèi)皮素（ET）抗體結(jié)合，形成抗體-抗原-酶標抗體復(fù)合物，經(jīng)過徹底洗滌后加底物TMB顯色。TMB在HRP酶的催化下轉(zhuǎn)化成藍色，并在酸的作用下轉(zhuǎn)化成Z終的黃色。顏色的深淺和樣品中的內(nèi)皮素（ET）呈正相關(guān)。用酶標儀在450nm波長下測定吸光度（OD值），通過標準曲線計算樣品中大鼠內(nèi)皮素（ET）濃度。人c-fosELISA試劑盒標本要求1．標本采集后盡早進行提取，提取按相關(guān)文獻進行，提取后應(yīng)盡快進行實驗。若不能馬上進行試驗，可將標本放于-20℃保存，但應(yīng)避免反復(fù)凍融2．不能檢測含NaN3的樣品，因NaN3YZ辣根過氧化物酶的（HRP）活性。操作步驟1.標準品的稀釋：本試劑盒提供原倍標準品一支，用戶可按照下列圖表在小試管中進行稀釋。120μg/L5號標準品150μl的原倍標準品加入150μl標準品稀釋液60μg/L4號標準品150μl的5號標準品加入150μl標準品稀釋液30μg/L3號標準品150μl的4號標準品加入150μl標準品稀釋液15μg/L2號標準品150μl的3號標準品加入150μl標準品稀釋液7.5μg/L1號標準品150μl的2號標準品加入150μl標準品稀釋液2.加樣：分別設(shè)空白孔（空白對照孔不加樣品及酶標試劑，其余各步操作相同）、標準孔、待測樣品孔。在酶標包被板上標準品準確加樣50μl，待測樣品孔中先加樣品稀釋液40μl，然后再加待測樣品10μl（樣品Z終稀釋度為5倍）。加樣將樣品加于酶標板孔底部，盡量不觸及孔壁，輕輕晃動混勻。3.溫育：用封板膜封板后置37℃溫育30分鐘。4.配液：將30倍濃縮洗滌液用蒸餾水30倍稀釋后備用5.洗滌：小心揭掉封板膜，棄去液體，甩干，每孔加滿洗滌液，靜置30秒后棄去，如此重復(fù)5次，拍干。6.加酶：每孔加入酶標試劑50μl，空白孔除外。7.溫育：操作同3。8.洗滌：操作同5。9.顯色：每孔先加入顯色劑A50μl，再加入顯色劑B50μl，輕輕震蕩混勻，37℃避光顯色15分鐘.10.終止：每孔加終止液50μl，終止反應(yīng)（此時藍色立轉(zhuǎn)黃色）。11.測定：以空白空調(diào)零，450nm波長依序測量各孔的吸光度（OD值）。測定應(yīng)在加終止液后15分鐘以內(nèi)進行。人c-fosELISA試劑盒注意事項1．試劑盒從冷藏環(huán)境中取出應(yīng)在室溫平衡15-30分鐘后方可使用，酶標包被板開封后如未用完，板條應(yīng)裝入密封袋中保存。2．濃洗滌液可能會有結(jié)晶析出，稀釋時可在水浴中加溫助溶，洗滌時不影響結(jié)果。3．各步加樣均應(yīng)使用加樣器，并經(jīng)常校對其準確性，以避免試驗誤差。一次加樣時間**控制在5分鐘內(nèi)，如標本數(shù)量多，推薦使用排槍加樣。4．請每次測定的同時做標準曲線，**做復(fù)孔。如標本中待測物質(zhì)含量過高（樣本OD值大于標準品孔**孔的OD值），請先用樣品稀釋液稀釋一定倍數(shù)（n倍）后再測定，計算時請Z后乘以總稀釋倍數(shù)（×n×5）。5．封板膜只限一次性使用，以避免交叉污染。6．底物請避光保存。7．嚴格按照說明書的操作進行，試驗結(jié)果判定必須以酶標儀讀數(shù)為準.8．所有樣品，洗滌液和各種廢棄物都應(yīng)按傳染物處理。9．本試劑不同批號組分不得混用。10.如與英文說明書有異，以英文說明書為準。保存條件及有效期1．試劑盒保存：；2-8℃。2．有效期：6個月[詳細]

  2018-09-19 10:00

  產(chǎn)品樣冊

  |

  • 
  • 
  • 
  • 

• 人一氧化氮(NO)ELISA試劑盒

  人一氧化氮(NO)ELISA試劑盒[詳細]

  2024-09-25 17:06

  操作手冊

  |

  • 
  • 
  • 
  • 

• 人preptin ELISA試劑盒

  人preptin ELISA試劑盒[詳細]

  2024-09-29 06:08

  報價單

  |

  • 
  • 
  • 
  • 

• 人ELISA試劑盒,TPA ELISA Kit

  96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人組織多肽抗原（TPA）ELISA試劑盒相關(guān)產(chǎn)品：96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人組織多肽抗原（TPA）ELISA試劑盒IL-10,小鼠白介素-10Elisa試劑盒磺胺二甲基惡唑,標準品JLJL200712人Elisa試劑盒,人ProteinCElisa試劑盒，HumanProteinCELISA試劑盒CAS號:108-69-0,3,5-二,98%人Elisa試劑盒,人CD30Elisa試劑盒，HumanClusterofdifferentiation30,CD30ELISA試劑盒CAS:893-36-7,鹽酸-L-白氨酰-2-萘胺/L-亮氨酰-2-萘胺鹽酸鹽/L-白氨酰-β-萘胺鹽酸鹽/鹽酸-L-亮氨酰-2-萘胺/L（+）-亮氨酰-2-萘基鹽酸氨/L-Leucyl-2-naphthylamidehydrochloride,BR,98%,1克,避光,-20℃96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人組織多肽抗原（TPA）ELISA試劑盒Humanbacterialvaginosis,BVELISA試劑盒人（BV）kit說明書,細菌性陰道病Elisa試劑盒CXCR3ELISAKit,大鼠CXC趨化因子受體3Elisa檢測試劑盒蒙花苷,標準品,含量測定,20mg,常溫,避光PorcineapoproteinA1,apo-A1ELISA試劑盒豬（apo-A1）kit說明書,載脂蛋白A1Elisa試劑盒大鼠淋巴細胞因子ELISA試劑盒HumanhepatitisBvirusXinteractingprotein,HBXIPELISAKit人異常凝血酶原（APT）ELISA試劑盒HumanAbnormalprothrombin,APTELISA試劑盒草烏甲素,標準品,含量測定,50mg,常溫,避光96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人組織多肽抗原（TPA）ELISA試劑盒GRP1957,營養(yǎng)肉湯,供一般細菌培養(yǎng)、轉(zhuǎn)種和增菌用,250g小鼠生長激素釋放多肽（GHRP）ELISA試劑盒HumanMotilin,MTLELISAKitCAS號:4767-3-7,2,2-雙（羥甲基）丙酸,98%[詳細]

  2018-10-23 10:31

  產(chǎn)品樣冊

  |

  • 
  • 
  • 
  • 

• Elisa試劑盒：人D-乳酸鹽Elisa試劑盒說明書

  Elisa試劑盒：人D-乳酸鹽Elisa試劑盒說明書人血清，血漿，細胞上清及相關(guān)液體樣本中D-乳酸鹽（D-lactate）的含量。Elisa試劑盒：人D-乳酸鹽Elisa試劑盒說明書用純化的D-乳酸鹽（D-lactate）抗體包被微孔板，制成固相抗體，往包被單抗的微孔中依次加入D-乳酸鹽（D-lactate），再與HRP標記的D-乳酸鹽（D-lactate）抗體結(jié)合，形成抗體-抗原-酶標抗體復(fù)合物，經(jīng)過徹底洗滌后加底物TMB顯色。Elisa試劑盒：人D-乳酸鹽Elisa試劑盒說明書濃縮洗滌液（20ml×20倍）×1瓶（20ml×30倍）×1瓶2-8℃保存Elisa試劑盒：人D-乳酸鹽Elisa試劑盒說明書人D-乳酸鹽（D-lactate）酶聯(lián)免疫分析（ELISA）試劑盒使用說明書本試劑僅供研究使用目的：本試劑盒用于測定人血清，血漿，細胞上清及相關(guān)液體樣本中D-乳酸鹽（D-lactate）的含量。實驗原理：本試劑盒應(yīng)用雙抗體夾心法測定標本中人D-乳酸鹽（D-lactate）水平。用純化的D-乳酸鹽（D-lactate）抗體包被微孔板，制成固相抗體，往包被單抗的微孔中依次加入D-乳酸鹽（D-lactate），再與HRP標記的D-乳酸鹽（D-lactate）抗體結(jié)合，形成抗體-抗原-酶標抗體復(fù)合物，經(jīng)過徹底洗滌后加底物TMB顯色。TMB在HRP酶的催化下轉(zhuǎn)化成藍色，并在酸的作用下轉(zhuǎn)化成Z終的黃色。顏色的深淺和樣品中的D-乳酸鹽（D-lactate）呈正相關(guān)。用酶標儀在450nm波長下測定吸光度（OD值），通過標準曲線計算樣品中人D-乳酸鹽（D-lactate）濃度。試劑盒組成：試劑盒組成48孔配置96孔配置保存說明書1份1份封板膜2片（48）2片（96）密封袋1個1個酶標包被板1×481×962-8℃保存標準品：450ng/ml0.5ml×1瓶0.5ml×1瓶2-8℃保存標準品稀釋液1.5ml×1瓶1.5ml×1瓶2-8℃保存酶標試劑3ml×1瓶6ml×1瓶2-8℃保存樣品稀釋液3ml×1瓶6ml×1瓶2-8℃保存顯色劑A液3ml×1瓶6ml×1瓶2-8℃保存顯色劑B液3ml×1瓶6ml×1瓶2-8℃保存終止液3ml×1瓶6ml×1瓶2-8℃保存濃縮洗滌液（20ml×20倍）×1瓶（20ml×30倍）×1瓶2-8℃保存樣本處理及要求：1.血清：室溫血液自然凝固10-20分鐘，離心20分鐘左右（2000-3000轉(zhuǎn)/分）。仔細收集上清，保存過程中如出現(xiàn)沉淀，應(yīng)再次離心。2.血漿：應(yīng)根據(jù)標本的要求選擇EDTA或檸檬酸鈉作為抗凝劑，混合10-20分鐘后，離心20分鐘左右（2000-3000轉(zhuǎn)/分）。仔細收集上清，保存過程中如有沉淀形成，應(yīng)該再次離心。3.尿液：用無菌管收集，離心20分鐘左右（2000-3000轉(zhuǎn)/分）。仔細收集上清，保存過程中如有沉淀形成，應(yīng)再次離心。胸腹水、腦脊液參照實行。4.細胞培養(yǎng)上清：檢測分泌性的成份時，用無菌管收集。離心20分鐘左右（2000-3000轉(zhuǎn)/分）。仔細收集上清。檢測細胞內(nèi)的成份時，用PBS（PH7.2-7.4）稀釋細胞懸液，細胞濃度達到100萬/ml左右。通過反復(fù)凍融，以使細胞破壞并放出細胞內(nèi)成份。離心20分鐘左右（2000-3000轉(zhuǎn)/分）。仔細收集上清。保存過程中如有沉淀形成，應(yīng)再次離心。5.組織標本：切割標本后，稱取重量。加入一定量的PBS，PH7.4。用液氮迅速冷凍保存?zhèn)溆谩吮救诨笕匀槐３?-8℃的溫度。加入一定量的PBS（PH7.4），用手工或勻漿器將標本勻漿充分。離心20分鐘左右（2000-3000轉(zhuǎn)/分）。仔細收集上清。分裝后一份待檢測，其余冷凍備用。6.標本采集后盡早進行提取，提取按相關(guān)文獻進行，提取后應(yīng)盡快進行實驗。若不能馬上進行試驗，可將標本放于-20℃保存，但應(yīng)避免反復(fù)凍融.7.不能檢測含NaN3的樣品，因NaN3YZ辣根過氧化物酶的（HRP）活性。操作步驟標準品的稀釋與加樣：在酶標包被板上設(shè)標準品孔10孔，在**、第二孔中分別加標準品100μl，然后在**、第二孔中加標準品稀釋液50μl，混勻；然后從**孔、第二孔中各取100μl分別加到第三孔和第四孔，再在第三、第四孔分別加標準品稀釋液50μl，混勻；然后在第三孔和第四孔中先各取50μl棄掉，再各取50μl分別加到第五、第六孔中，再在第五、第六孔中分別加標準品稀釋液50ul，混勻；混勻后從第五、第六孔中各取50μl分別加到第七、第八孔中，再在第七、第八孔中分別加標準品稀釋液50μl，混勻后從第七、第八孔中分別取50μl加到第九、第十孔中，再在第九第十孔分別加標準品稀釋液50μl，混勻后從第九第十孔中各取50μl棄掉。（稀釋后各孔加樣量都為50μl，濃度分別為300ng/ml，200ng/ml，100ng/ml，50ng/ml，25ng/ml）。加樣：分別設(shè)空白孔（空白對照孔不加樣品及酶標試劑，其余各步操作相同）、待測樣品孔。在酶標包被板上待測樣品孔中先加樣品稀釋液40μl，然后再加待測樣品10μl（樣品Z終稀釋度為5倍）。加樣將樣品加于酶標板孔底部，盡量不觸及孔壁，輕輕晃動混勻。溫育：用封板膜封板后置37℃溫育30分鐘。配液：將30（48T的20倍）倍濃縮洗滌液用蒸餾水30（48T的20倍）倍稀釋后備用。洗滌：小心揭掉封板膜，棄去液體，甩干，每孔加滿洗滌液，靜置30秒后棄去，如此重復(fù)5次，拍干。加酶：每孔加入酶標試劑50μl，空白孔除外。溫育：操作同3。洗滌：操作同5。顯色：每孔先加入顯色劑A50μl，再加入顯色劑B50μl，輕輕震蕩混勻，37℃避光顯色15分鐘.終止：每孔加終止液50μl，終止反應(yīng)（此時藍色立轉(zhuǎn)黃色）。測定：以空白空調(diào)零，450nm波長依序測量各孔的吸光度（OD值）。測定應(yīng)在加終止液后15分鐘以內(nèi)進行。注意事項：試劑盒從冷藏環(huán)境中取出應(yīng)在室溫平衡15-30分鐘后方可使用，酶標包被板開封后如未用完，板條應(yīng)裝入密封袋中保存。濃洗滌液可能會有結(jié)晶析出，稀釋時可在水浴中加溫助溶，洗滌時不影響結(jié)果。各步加樣均應(yīng)使用加樣器，并經(jīng)常校對其準確性，以避免試驗誤差。一次加樣時間**控制在5分鐘內(nèi)，如標本數(shù)量多，推薦使用排槍加樣。請每次測定的同時做標準曲線，**做復(fù)孔。如標本中待測物質(zhì)含量過高（樣本OD值大于標準品孔**孔的OD值），請先用樣品稀釋液稀釋一定倍數(shù)（n倍）后再測定，計算時請Z后乘以總稀釋倍數(shù)（×n×5）。封板膜只限一次性使用，以避免交叉污染。底物請避光保存。嚴格按照說明書的操作進行，試驗結(jié)果判定必須以酶標儀讀數(shù)為準.所有樣品，洗滌液和各種廢棄物都應(yīng)按傳染物處理。本試劑不同批號組分不得混用。10.如與英文說明書有異，以英文說明書為準。計算：以標準物的濃度為橫坐標，OD值為縱坐標，在坐標紙上繪出標準曲線，根據(jù)樣品的OD值由標準曲線查出相應(yīng)的濃度；再乘以稀釋倍數(shù)；或用標準物的濃度與OD值計算出標準曲線的直線回歸方程式，將樣品的OD值代入方程式，計算出樣品濃度，再乘以稀釋倍數(shù)，即為樣品的實際濃度。（此圖僅供參考）試劑盒性能：1.樣品線性回歸與預(yù)期濃度相關(guān)系數(shù)R值為0.92以上。2.批內(nèi)與批見應(yīng)分別小于9%和15%檢測范圍：8ng/ml-400ng/ml保存條件及有效期：1.試劑盒保存：；2-8℃。2．有效期：6個月[詳細]

  2018-10-23 10:31

  產(chǎn)品樣冊

  |

  • 
  • 
  • 
  • 

• 人巨噬細胞集落刺激因子(M-CSF)ELISA試劑盒

  人巨噬細胞集落刺激因子(M-CSF)ELISA試劑盒[詳細]

  2013-12-06 00:00

  課件

  |

  • 
  • 
  • 
  •