BioCheck新产品BC-1303 说明书
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世界ding级实验材料供应商BioCheck正式授权上海起发为其ZG代理，BioCheck在一直是行业的标杆,一直为广大科研客户提供Z为优质的产品和服务,上海起发一直秉承为ZG科研客户带来**的产品，**的服务，签约BioCheck就是为了给广大科研客户带来更加完善的产品和服务,您的满意将是我们Zda的收获BioCheckZG代理,BioCheck上海代理，BioCheck北京代理，BioCheck广东代理，BioCheck江苏代理BioCheck湖北代理,BioCheck天津,BioCheck黑龙江代理,BioCheck内蒙古代理,BioCheck吉林代理,BioCheck福建代理,BioCheck江苏代理,BioCheck浙江代理,BioCheck四川代理BIOCHECK公司由创始人Dr.JohnChen,Medix创立,是一家抓也提供肿瘤标志物，心肌标志物，激素类Z高性价比的抗体的公司。简要原理利用竞争酶联免疫方法，预先在微孔中包被羊抗兔抗体，实验时先后加入氯霉素标准品或待测样本，氯霉素酶标抗原和兔抗氯霉素抗体。经过室温温育，反应液中的兔抗氯霉素抗体与微孔板上的羊抗兔抗体结合，待测样品中的抗原与氯霉素酶标抗原竞争微孔板上的兔抗氯霉素抗体。洗涤后，没有与抗体结合的待测样品中的抗原或酶标抗原被洗去，再加入反应底物，结合的酶标抗原的酶将底物转化为蓝色产物，加入终止液后颜色由蓝色变为黄色。反应完成后，样品中氯霉素含量越多，反应呈色就越浅；反之，样品中氯霉素含量越少，则呈色越深。利用标准曲线可计算出样品中氯霉素含量。技术参数检测蜂蜜、蛋类、牛奶、奶粉、水产品、动物组织(肌肉、肝脏等)、饲料、血清、血浆及尿液样本中存在的氯霉素，定量限可达0.025ppb。No.样品检测下限1蜂蜜0.02ppb(0.02ng/g)2蛋类0.02ppb(0.02ng/g)3牛奶0.002ppb(0.002ng/ml)4奶粉0.012ppb(0.012ng/g)5虾，鱼及肉类0.08ppb(0.08ng/g)6饲料0.08ppb(0.08ng/g)7血清/血浆0.02ppb(0.02ng/ml)8尿液0.04ppb(0.04ng/ml)交叉反应名称百分比氯霉素碱0.4%甲基氯霉素<0.04%回收率No.样品回收率1蜂蜜70%~110%2牛奶90%~130%3蛋，虾，鱼及肉类95%~120%4饲料95%~120%5血清/血浆90%~120%6尿液1**%~130%PD-L1ENZYMEIMMUNOASSAYTESTKITCatalogNumber:BC-1303EnzymeImmunoassayfortheQuantitativeDeterminationofPD-L1ConcentrationinHumanSerumandPlasmaFORRESEARCHUSEONLYNotforuseindiagnosticproceduresINTRODUCTIONProgrammeddeathreceptor-ligand1(PD-L1,B7-H1,CD274)isabiomarkerfromtheB7familythatisexpressedonavarietyofcellsandupregulatedinresponsetopro-inflammatorycytokinessuchasinterferongamma1,2,3.PD-L1interactswithPD-1,aco-receptorexpressedbyexhaustedTcells,toencourageanimmunosuppressivetumormicroenvironmentbydecreasingTcellreceptormediatedproliferationandcytokineproduction4,5.ThePD-1/PD-L1interactionfunctionsasanimmunecheckpointinaprocessknownasimmunoeditingwherethehostimmunesystemeliminateshighlyimmunogenictumorswhileallowinglessimmunogenictumortoevadeit2,6.Multiplesolidtumortypesincludingmelanoma,renalcellcarcinoma,non-smallcelllungcancer,ovarian,andcolorectalcancerutilizethisPD-1/PD-L1immunoeditingmechanism2.CurrenttreatmenthasfocusedonblockingthePD-1/PD-L1interactiontoreducetumorevasionbyinhibitingPD-1,withnewfocusonPD-L1aswell1,2.PRINCIPLEOFTHEASSAYThePD-L1ELISAisbasedontheprincipleofasolidphaseenzyme-linkedimmunosorbentassay.TheassaysystemutilizesauniquemonoclonalantibodypairdirectedagainstadistinctantigenicdeterminantonthePD-L1molecule.Onemousemonoclonalanti-PD-L1antibodyisusedforsolidphaseimmobilization(onthemicrotiterwells).Anothermousemonoclonalanti-PD-L1antibodyconjugatedtohorseradishperoxidase(HRP)isintheenzymeconjugatesolution.Thetestsamplesareallowedtoreactsequentiallywiththetwoantibodies,resultinginthePD-L1moleculestobesandwichedbetweenthesolidphaseandenzyme-linkedantibodies.Aftertwoseparate60-minuteincubationstepsatroomtemperature,thewellsarerinsedwithWashBuffertoremoveunboundlabeledantibodies.TMBReagentisaddedandincubatedfor30minutesunderdarkconditions,resultinginthedevelopmentofabluecolor.ThecolordevelopmentisstoppedwiththeadditionofStopSolution,changingthecolortoyellow.TheconcentrationofPD-L1isdirectlyproportionaltothecolorintensityofthetestsamples.Absorbanceismeasuredspectrophotometricallyat450nm.REAGENTSANDMATERIALSPROVIDEDAntibody-CoatedWells(1plate,96wells)Microtiterwellscoatedwithmousemonoclonalanti-PD-L120ng/mlPD-L1Standard(0.5mL/vial)20ng/mLPD-L1inphosphatebuffer-BSAsolutionwithpreservatives3.StandardandSampleDiluent(30mL/bottle,1bottle)Containsphosphatebuffer-BSAsolutionwithpreservativesEnzymeConjugateReagent(12mL/vial,1vial)Containsmousemonoclonalanti-PD-L1conjugatedtohorseradishperoxidase20XWashBuffer(50mL/bottle,1bottle)PhosphatebufferwithdetergentsTMBReagent(11mL/bottle,1bottle)Containsone-stepTMBsolutionStopSolution(11mL/bottle,1bottle)Containsdilutedhydrochloricacid(1NHCl)STORAGECONDITIONSStoretheunopenedkitat2-8°Cuponreceiptandwhenitisnotinuse,untiltheexpirationshownonthekitlabel.Refertothepackagelabelfortheexpirationdate.Keepmicrotiterplateinasealedbagwithdesiccanttominimizeexposuretodampair.REAGENTPREPARATIONAllreagents首ldbeallowedtoreachroomtemperature(18-25°C)beforeuse.Foreachtestrun,prepareafreshstandardset.Dilute20ng/mLstandardto5ng/mL.Preparetwo-foldserialdilutionsofthe5ng/mLStandardwithStandard/SampleDiluent:5ng/mL:0.15mLof20ng/mL+0.45mLofStandard/SampleDiluent2.5ng/mL:0.25mLof5ng/mL+0.25mLofStandard/SampleDiluent1.25ng/mL:0.25mLof2.5ng/ml+0.25mLofStandard/SampleDiluent0.625ng/mL:0.25mLof1.25ng/mL+0.25mLofStandard/SampleDiluent0.313ng/mL:0.25mLof0.625ng/mL+0.25mLofStandard/SampleDiluent0.156ng/mL:0.25mLof0.313ng/mL+0.25mLofStandardDiluent0.078ng/mL:0.25mLof0.156ng/mL+0.25mLofStandardDiluent0ng/mL:0.25mLofStandardDiluentPatientsamplesneedtobediluted4-foldpriortouse.Prepareaseriesofsmalltubes(i.e.,1.5mLmicrocentrifugetubes)andmix60Lofserumwith180LStandard/SampleDiluent.WorkingWashBuffer:Preparationof1XWashBufferfrom20XStock.Add50mLof20XWashBufferStockto950mLofDIH2O.TheWorkingWashBufferisstableat2-8°Cfor30days.NOTE:Anycrystalsthatmaybepresentduetohighsaltconcentrationmustberedissolvedatroomtemperaturebeforemakingthedilution.ASSAYPROCEDUREPrepareStandards.SeeReagentPreparation.Dilutesamples1:4dilution.SeeReagentPreparation.Securethedesirednumberofcoatedwellsintheholder.Dispense100mLofPD-L1standards,andDILUTEDspecimensintotheappropriatewells.Incubatefor60minutesatroomtemperature(18-25°C).Removeincubationmixturebyflickingplatecontentsintoawastecontainer.Rinseandflickthemicrotiterwells5timeswith300mLWorkingWashBuffer.Strikethewellsontoabsorbentpaperorpapertowelstoremoveallresidualwaterdroplets.Dispense100mLofPD-L1WorkingEnzymeConjugateReagentintoeachwell.Incubatefor60minutesatroomtemperature(18-25°C).Removeincubationmixturebyflickingplatecontentsintoawastecontainer.Rinseandflickthemicrotiterwells5timeswith300uLWorkingWashBuffer.Strikethewellsontoabsorbentpaperorpapertowelstoremoveallresidualwaterdroplets.Dispense100mLTMBsolutionintoeachwell.Incubatefor30minutesatroomtemperature(18-25°C).Stopthereactionbyadding100mLofStopSolutionintoeachwell.Gentlymixfor30seconds.Itisimportanttomakesurethatallthebluecolorchangestoyellowcolorcompletely.Readabsorbanceat450nmwithamicrotiterwellreaderwithin15minutes.CALCULATIONOFRESULTSCalculatethemeanabsorbancevalue(OD450)foreachsetofreferencestandards,controlsandsamples.Constructastandardcurvebyplottingthemeanabsorbanceobtainedforeachreferencestandardagainstitsconcentrationinng/mlongraphpaper,withabsorbanceonthevertical(y)axisandconcentrationonthehorizontal(x)axis.ThecorrespondingconcentrationofPD-L1(ng/mL)canbedeterminedfromthestandardcurveusingthemeanabsorbancevalueforeachsample.Dependingonexperienceand/ortheavailabilityofcomputercapability,othermethodsofdatareductionmaybeemployed.Theobtainedvaluesofthesamples首ldbemultipliedbythedilutionfactorof4toobtainPD-L1resultsinng/ml.EXAMPLEOFSTANDARDCURVEResultsofatypicalstandardrunwithabsorbencyreadingsat450nmshownontheYaxisagainstPD-L1concentrationsshownontheXaxis.NOTE:Thisstandardcurveisforthepurposeofillustrationonly,and首ldnotbeusedtocalculateunknowns.Eachlaboratorymustgenerateitsowndataandstandardcurveineachexperiment.PD-L1Absorbance(ng/ml)(450nm)00.0420.0780.1410.1560.2360.3130.4150.6250.7261.251.2832.52.10053.053nm3.53(4502.52Absorbance1.510.50012345PD-L1Conc.(ng/ml)PERFORMANCECHARACTERISTICSSensitivityTheminimumdetectableconcentrationofthePD-L1ELISAassayasmeasuredby2SDfromthemeanofazerostandardisestimatedtobe0.02ng/ml.PrecisionIntra-AssayPrecisionWithin-runprecisionwasdeterminedbyreplicatedeterminationsofthreedifferentsamplesinoneassay.Within-assayvariabilityisshownbelow:Sample123#Replicates242424MeanPD-L1(ng/mL)1.53.07.4S.D.0.040.050.18C.V.(%)2.4%1.6%2.4%2Inter-AssayPrecisionBetween-runprecisionwasdeterminedbyreplicatemeasurementsofthreedifferentsamplesoveraseriesofindividuallycalibratedassays.Between-assayvariabilityisshownbelow:Sample123#Replicates202020MeanPD-L1(ng/mL)1.63.07.4S.D.0.040.120.26C.V.(%)2.5%4.1%3.5%RecoveryandLinearityStudiesRecoverySampleswerespikedwithknownPD-L1levelsandassayedinduplicate.Themeanrecoverywas90%.SampleEXPECTEDOBSERVED%RECOVERY[PD-L1][PD-L1](ng/mL)(ng/mL)121.890%254.692%3108.989%LinearityThreesampleswereseriallydilutedtodeterminelinearity.Themeanrecoverywas110.3%.#DilutionExpectedObservedConc.(ng/ml)Conc.(ng/ml)%Expected1.1:41.61.6N/A1:81.7106.3%1:161.7106.3%1:321.8112.5%Mean=108.4%2.1:43.73.7N/A1:84.0108.1%1:164.0108.1%1:324.1110.8%Mean=109.0%3.1:47.47.4N/A1:88.3112.2%1:168.6116.2%1:328.3112.2%Mean=113.5%REFERENCESHerbst,RoyS.,etal."Predictivecorrelatesofresponsetotheanti-PD-L1antibodyMPDL3280Aincancerpatients."Nature515.7528(2014):563.Patel,SandipPravin,andRazelleKurzrock."PD-L1expressionasapredictivebiomarkerincancerimmunotherapy."Molecularcancertherapeutics14.4(2015):847-856.Topalian,SuzanneL.,CharlesG.Drake,andDrewM.Pardoll."TargetingthePD-1/B7-H1(PD-L1)pathwaytoactivateanti-tumorimmunity."Currentopinioninimmunology24.2(2012):207-212.Blank,Christian,andAndreasMackensen."ContributionofthePD-L1/PD-1pathwaytoT-cellexhaustion:anupdateonimplicationsforchronicinfectionsandtumorevasion."Cancerimmunology,immunotherapy56.5(2007):739-745.Barber,DanielL.,etal."RestoringfunctioninexhaustedCD8Tcellsduringchronicviralinfection."Nature439.7077(2006):682.Teng,MicheleWL,etal."ClassifyingcancersbasedonT-cellinfiltrationandPD-L1."Cancerresearch75.11(2015):2139-2145.部分Zxin产品如下：货号品名Tests/Kit规格品牌70748MouseMonoclonalanti-humanPD-1(Capture)Purified0.1/0.5/1mgBioCheck70749MouseMonoclonalanti-humanPD-1(Capture)Purified0.1/0.5/1mgBioCheck70750MouseMonoclonalanti-humanPD-1(Detection)Purified0.1/0.5/1mgBioCheck70751MouseMonoclonalanti-humanPD-L1(Capture)Purified0.1/0.5/1mgBioCheck70752MouseMonoclonalanti-humanPD-L1(Capture)Purified0.1/0.5/1mgBioCheck70753MouseMonoclonalanti-humanPD-L1(Detection)Purified0.1/0.5/1mgBioCheck70745Mousemonoclonalanti-humanLp-PLA2(Capture)Purified0.5mgBioCheck70746Mousemonoclonalanti-humanLp-PLA2(Detection)Purified0.5mgBioCheck70747Mousemonoclonalanti-humanLp-PLA2(Detection)Purified0.5mgBioCheck我们公司Zda优势是强大的采购，1：基本什么都能进口，血清，抗体，耗材，还有部分限制进口的，2：货品全，现经营过700多个品牌，基本所有生物试剂耗材都可以进口，特别是冷偏的产品那就更有优势，3：提供加急服务，一般1-2周到货，超过时限加急费全免4：价格公道，绝大部分价格有优势，当然不能保证1**%产品都是价格Zdi,因为价格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