红细胞生成素（EPO）Elisa试剂盒下载资料
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FORRESEARCHUSEONLYRatErythropoietinDrugNamesGenericName：RatErythropoietin（EPO）ELISAKit.PurposeThiskitallowsforthedeterminationofEPOconcentrationsinRatserum,bloodplasma,andotherbiologicalfluids.PrincipleoftheassayThekitassayRatEPOlevelinthesample，usePurifiedRatEPOantibodytocoatmicrotiterplatewells,makesolid-phaseantibody,thenaddEPOtowells,CombinedEPOantibodywhichWithHRPlabeled,becomeantibody-antigen-enzyme-antibodycomplex,afterwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolorAtHRPenzyme-catalyzed,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofEPOinthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.MaterialsprovidedwiththekitMaterialsprovidedwiththekit48determinations96determinationsStorageUsermanual11Closureplatemembrane22Sealedbags11Microelisastripplate112-8℃Standard：27IU/L0.5ml×1bottle0.5ml×1bottle2-8℃Standarddiluent1.5ml×1bottle1.5ml×1bottle2-8℃HRP-Conjugatereagent3ml×1bottle6ml×1bottle2-8℃Samplediluent3ml×1bottle6ml×1bottle2-8℃ChromogenSolutionA3ml×1bottle6ml×1bottle2-8℃ChromogenSolutionB3ml×1bottle6ml×1bottle2-8℃StopSolution3ml×1bottle6ml×1bottle2-8℃washsolution（20ml×20fold）×1bottle（20ml×30fold）×1bottle2-8℃Specimenrequirements1.serum-coagulationatroomtemperature10-20mins，centrifugation20-minatthespeedof2000-3000r.p.m.removesupernatant,Ifprecipitationappeared,Centrifugalagain.2.plasma-usesuitedEDTAorcitrateplasmaasananticoagulant,mix10-20mins,centrifugation20-minatthespeedof2000-3000r.p.m.removesupernatant,Ifprecipitationappeared,Centrifugalagain.3.Urine-collectsueasterilecontainer,centrifugation20-minatthespeedof2000-3000r.p.m.removesupernatant,Ifprecipitationappeared,Centrifugalagain.TheOperationofHydrothoraxandcerebrospinalfluidReferencetoit.4.cellculturesupernatant-detectsecretorycomponents,collectsueasterilecontainer,centrifugation20-minatthespeedof2000-3000r.p.m.removesupernatant,detectthecompositionofcells,DilutcellsuspensionwithPBS（PH7.2-7.4）,Cellconcentrationreached1million/ml,repeatedfreeze-thawcycles,damagecellsandreleaseofintracellularcomponents,centrifugation20-minatthespeedof2000-3000r.p.m.removesupernatant,Ifprecipitationappeared,Centrifugalagain.5.Tissuesamples-Aftercuttingsamples,checktheweight,addPBS（PH7.2-7.4）,Rapidlyfrozenwithliquidnitrogen,maintainsamplesat2-8℃aftermelting,addPBS（PH7.4）,HomogenizedbyhandorGrinders,centrifugation20-minatthespeedof2000-3000r.p.m.removesupernatant.6.extractassoonaspossibleafterSpecimencollection,andaccordingtotherelevantliterature,and首ldbeexperimentassoonaspossibleaftertheextraction.Ifitcan’t,specimencanbekeptin-20℃topreserve,Avoidrepeatedfreeze-thawcycles.7.Can’tdetectthesamplewhichcontainNaN3,becauseNaN3inhibitsHRPactive.Assayprocedure1.DiluteandaddsampletoStandard:set10StandardwellsontheELISAplatescoated,addStandard100μltothefirstandthesecondwell,thenaddStandarddilution50μltothefirstandthesecondwell,mix;takeout100μlformthefirstandthesecondwellthenaddittothethirdandtheforthwellseparately.thenaddStandarddilution50μltothethirdandtheforthwell,mix;thentakeout50μlfromthethirdandtheforthwelldiscard,add50μltothefifthandthesixthwell,thenaddStandarddilution50μltothefifthandthesixthwell,mix;takeout50μlfromthefifthandthesixthwellandaddtotheseventhandtheeighthwell,thenaddStandarddilution50μltotheseventhandtheeighthwell,mix;takeout50μlfromtheseventhandtheeighthwellandaddtotheninthandthetenthwell,addStandarddilution50μltotheninthandthetenthwell,mix,takeout50μlfromtheninthandthetenthwelldiscard（addSample50μltoeachwellafterDiluting,（density:18IU/L，12IU/L，6IU/L，3IU/L，1.5IU/L）2.addsample：Setblankwellsseparately（blankcomparisonwellsdon’taddsampleandHRP-Conjugatereagent,othereachstepoperationissame）.testingsamplewell.addSampledilution40μltotestingsamplewell,thenaddtestingsample10μl（samplefinaldilutionis5-fold）,addsampletowells,don’ttouchthewellwallasfaraspossible,andGentlymix.3.Incubate:AfterclosingplatewithClosureplatemembrane,incubatefor30minat37℃.4.Configurateliquid:30-fold（or20-fold）washsolutiondiluted30-fold（or20-fold）withdistilledwaterandreserve.5.washing：UncoverClosureplatemembrane,discardLiquid,drybyswing,addwashingbuffertoeverywell,stillfor30sthendrain,repeat5times,drybypat.6.addenzyme：AddHRP-Conjugatereagent50μltoeachwell,exceptblankwell.7.incubate：Operationwith3.8.washing：Operationwith5.9.color：AddChromogenSolutionA50ulandChromogenSolutionBtoeachwell,evadethelightpreservationfor15minat37℃10.Stopthereaction：AddStopSolution50μltoeachwell,Stopthereaction（thebluecolorchangetoyellowcolor）.11.assay：takeblankwellaszero,Readabsorbanceat450nmafterAddingStopSolutionandwithin15min.Importantnotes1.Thekittakesoutfromtherefrigerationenvironment首ldbebalanced15-30minutesintheroomtemperature,ELISAplatescoatedifhasnotuseupafteropened,theplate首ldbestoredinSealedbag.2.washingbufferwillCrystallizationseparation,itcanbeheatedthewaterhelpsdissolvewhendilute.Washingdoesnotaffecttheresult.3.addSamplewithsamplerEachstep,Andproofreaditsaccuracyfrequently,avoidstheexperimentalerror.addsamplewithin5mins,ifthenumberofsampleismuch,recommendtouseVolley.4.ifthetestingmaterialcontentisexcessivelyhigher（ThesampleODisbiggerthanthefirststandardwell）,pleasediluteSample（n-fold）,Pleasediluenteandmultipliedbythedilutionfactor.（×n×5）.5.Closureplatemembraneonlylimitsthedisposableuse,toavoidcross-contamination.6.Thesubstrateevadethelightpreservation.7.Pleaseaccordingtouseinstructionstrictly,Thetestresultdeterminationmusttakethemicrotiterplatereaderasastandard.8.Allsamples,washingbufferandeachkindofreject首ldaccordingtoinfectivematerialprocess.9.Donotmixreagentswiththosefromotherlots.Takethestandarddensityasthehorizontal,theODvalueforthevertical,drawthestandardcurveongraphpaper,FindoutthecorrespondingdensityaccordingtothesampleODvaluebytheSamplecurve,multipliedbythedilutionmultiple,orcalculatethestraightlineregressionequationofthestandardcurvewiththestandarddensityandtheODvalue,withthesampleODvalueintheequation,calculatethesampledensity,multipliedbythedilutionfactor,theresultisthesampleactualdensity.CalculateThischartisforreferenceonlyAssayrange1IU/L20IU/LStorageandvalidity1．Storage：2-8℃.2．validity：sixmonths.上海广锐生物科技有限公司专业经销众多国际生命科学领域的知名品牌ELISA试剂盒，生物试剂，抗体，培养基，血清。为ZG科研试剂权威供应商之一，质量被全国各大院校科研机构认可，并指定为进口Elisa试剂盒/国产Elisa试剂盒长期供应商，作为战略合作伙伴。更多Elisa试剂盒、试剂优惠请登录本公司网站：http://www.shgrsw.com
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