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佐劑誘導(dǎo)的關(guān)節(jié)炎(Rat AIA)造模流程
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2018-10-20 10:00 1104閱讀次數(shù)
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小鼠關(guān)節(jié)炎(CIA)造膜流程(Chondrex)
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富氫水對關(guān)節(jié)炎的功效
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大鼠(Rat)低氧誘導(dǎo)因子1α(HIF-1α)ELISA檢測試劑盒說明書
- 本試劑僅供研究使用ELISA檢測試劑盒試驗(yàn)原理:HIF-1α試劑盒是固相夾心法酶聯(lián)免疫吸附實(shí)驗(yàn)(ELISA).已知HIF-1α濃度的標(biāo)準(zhǔn)品、未知濃度的樣品加入微孔酶標(biāo)板內(nèi)進(jìn)行檢測。先將HIF-1α和生物素標(biāo)記的抗體同時(shí)溫育。洗滌后,加入親和素標(biāo)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(shí)作用。產(chǎn)生顏色。顏色的深淺和樣品中HIF-1α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(biāo)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(biāo)準(zhǔn)品:800pg/ml1瓶(0.6ml)1瓶(0.3ml)按說明書進(jìn)行稀稀空白對照1瓶(1.0ml)1瓶(0.5ml)即用型標(biāo)準(zhǔn)品稀釋緩沖液1瓶(4.0ml)1瓶(2.0ml)即用型生物素標(biāo)記的抗HIF-1α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(jìn)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(biāo)本稀釋液1瓶(12ml)1瓶(6.0ml)即用型自備材料蒸餾水。加樣器:5ul、10ul、50ul、100ul、200ul、500ul、1000ul。振蕩器及磁力攪拌器等。安全性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。實(shí)驗(yàn)中不要吃喝、抽煙或使用化妝品。不要用嘴吸取試劑盒里的任何成份。ELISA檢測試劑盒操作注意事項(xiàng)試劑應(yīng)按標(biāo)簽說明書儲存,使用前恢復(fù)到室溫。稀稀過后的標(biāo)準(zhǔn)品應(yīng)丟棄,不可保存。實(shí)驗(yàn)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)。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。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(shí),避免使用帶金屬部分的加樣器。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。洗滌酶標(biāo)板時(shí)應(yīng)充分拍干,不要將吸水紙直接放入酶標(biāo)反應(yīng)孔中吸水。底物A應(yīng)揮發(fā),避免長時(shí)間打開蓋子。底物B對光敏感,避免長時(shí)間暴露于光下。避免用手接觸,有毒。實(shí)驗(yàn)完成后應(yīng)立即讀取OD值。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(shí)間一樣。按照說明書中標(biāo)明的時(shí)間、加液的量及順序進(jìn)行溫育操作。樣品收集、處理及保存方法血清-----操作過程中避免任何細(xì)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(xì)胞迅速小心地分離。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。細(xì)胞上清液---1000×g離心10分鐘去除顆粒和聚合物。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。試劑的準(zhǔn)備標(biāo)準(zhǔn)品:標(biāo)準(zhǔn)品的系列稀釋應(yīng)在實(shí)驗(yàn)時(shí)準(zhǔn)備,不能儲存。稀釋前將標(biāo)準(zhǔn)品振蕩混勻。稀釋比例按下表中進(jìn)行:800pg/ml(6號標(biāo)準(zhǔn)品)原倍濃度不用稀釋直接加入50ul。400pg/ml(5號標(biāo)準(zhǔn)品)100ul的原倍標(biāo)準(zhǔn)品加入100ul的標(biāo)準(zhǔn)品稀釋液200pg/ml(4號標(biāo)準(zhǔn)品)100ul的5號標(biāo)準(zhǔn)品加入100ul的標(biāo)準(zhǔn)品稀釋液100pg/ml(3號標(biāo)準(zhǔn)品)100ul的4號標(biāo)準(zhǔn)品加入100ul的標(biāo)準(zhǔn)品稀釋液50pg/ml(2號標(biāo)準(zhǔn)品)100ul的3號標(biāo)準(zhǔn)品加入100ul的標(biāo)準(zhǔn)品稀釋液25pg/ml(1號標(biāo)準(zhǔn)品)100ul的2號標(biāo)準(zhǔn)品加入100ul的標(biāo)準(zhǔn)品稀釋液0pg/ml(空白對照)原始濃度不用稀釋直接加入50ul。洗滌緩沖液(50×)的稀釋:蒸餾水50倍稀釋。操作步驟使用前,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(shí)加入大量的氣泡,產(chǎn)生加樣上的誤差。根據(jù)待測樣品數(shù)量加上標(biāo)準(zhǔn)品的數(shù)量決定所需的板條數(shù)。每個標(biāo)準(zhǔn)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(biāo)本用標(biāo)本稀釋液1:1稀釋后加入50ul于反應(yīng)孔內(nèi)。加入稀釋好后的標(biāo)準(zhǔn)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(biāo)記的抗體。蓋上膜板,輕輕振蕩混勻,37℃溫育1小時(shí)。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(jī)洗滌,洗滌次數(shù)增加一次。每孔加入60ul的親和鏈酶素-HRP,輕輕振蕩混勻,37℃溫育30分鐘。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(jī)洗滌,洗滌次數(shù)增加一次。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。取出酶標(biāo)板,迅速加入50ul終止液,加入終止液后應(yīng)立即測定結(jié)果。在450nm波長處測定各孔的OD值。建議使用的實(shí)驗(yàn)方案標(biāo)準(zhǔn)品濃度(pg/ml)A800800樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品B400400樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品C200200樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品D100100樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品E5050樣品樣品樣品樣品樣品樣品樣品樣品樣品樣品F2525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(biāo)準(zhǔn)品以上的結(jié)果為非線性的,根據(jù)此標(biāo)準(zhǔn)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(biāo)準(zhǔn)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(xì)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(biāo)儀上讀取各孔的OD值2、以吸光度OD值為縱坐標(biāo)(Y),相應(yīng)的HIF-1α標(biāo)準(zhǔn)品濃度為橫坐標(biāo)(X),做得相應(yīng)的曲線,樣品的HIF-1α含量可根據(jù)其OD值由標(biāo)準(zhǔn)曲線換算出相應(yīng)的濃度。3、ELISA檢測試劑盒檢測值范圍:0-800pg/ml4、敏感度:1.0pg/ml[詳細(xì)]
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2018-09-27 10:00
產(chǎn)品樣冊
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Rat myeloperoxidase
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RatmyeloperoxidaseFORRESEARCHUSEONLYDrugNamesGenericName:Ratmyeloperoxidase(MPO)ELISAKit.PurposeThiskitallowsforthedeterminationofMPOconcentrationsinRatserum,bloodplasma,andotherbiologicalfluids.PrincipleoftheassayThekitassayRatMPOlevelinthesample,usePurifiedRatMPOantibodytocoatmicrotiterplatewells,makesolid-phaseantibody,thenaddMPOtowells,CombinedMPOantibodywhichWithHRPlabeled,becomeantibody-antigen-enzyme-antibodycomplex,afterwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolorAtHRPenzyme-catalyzed,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofMPOinthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.MaterialsprovidedwiththekitMaterialsprovidedwiththekit48determinations96determinationsStorageUsermanual11Closureplatemembrane22Sealedbags11Microelisastripplate112-8℃Standard:180U/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-usesuitedEDTAorcitrateorasananticoagulant,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:120U/L,80U/L,40U/L,20U/L,10U/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.addsamplewithin5min,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.CalculateThischartisforreferenceonlyAssayrange8U/L-150U/LStorageandvalidity1.Storage:2-8℃.2.validity:sixmonths.[詳細(xì)]
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2018-10-23 10:31
產(chǎn)品樣冊
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大鼠非酒精性脂肪肝造模方法的改進(jìn)
- 大鼠非酒精性脂肪肝造模方法的改進(jìn)[詳細(xì)]
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2014-08-19 00:00
操作手冊
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聚酰胺尼龍鑄造模粉的粘度測量
- 聚酰胺尼龍鑄造模粉的粘度測量[詳細(xì)]
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2014-07-11 00:00
標(biāo)準(zhǔn)
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Rat LPS試劑盒
- RatLPS試劑盒RatLPSFORRESEARCHUSEONLYAssayrange:0.03Eu/L0.8Eu/L96determinationsPurposeThiskitallowsforthedeterminationofLPSconcentrationsinRatserum,cellculturesupernatesandotherbiologicalfluidsRatLPS試劑盒PrincipleoftheassayThekitassayRatLPSlevelinthesample,usePurifiedRatLPSantibodytocoatmicrotiterplatewells,makesolid-phaseantibody,thenaddLPStowells,CombinedLPSantibodywhichWithHRPlabeled,becomeantibody-antigen-enzyme-antibodycomplex,afterwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolorAtHRPenzyme-catalyzed,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofRatLPSinthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.RatLPS試劑盒Materialsprovidedwiththekit1washsolution20ml×1bottle7StopSolution6ml×1bottle2HRP-Conjugatereagent6ml×1bottle8Standard(1.6Eu/L)0.5ml×1bottle3Microelisastripplate12well×8strips9Standarddiluent1.5ml×1bottle4Samplediluent6ml×1bottle10Instruction15ChromogenSolutionA6ml×1bottle11ClosureplateRatLPS試劑盒membrane26ChromogenSolutionB6ml×1bottle12Sealedbags1Specimenrequirements1.extractassoonaspossibleafterSpecimencollection,andaccordingtotherelevant2literature,and首ldbeexperimentassoonaspossibleaftertheextraction.Ifitcan’t,specimencanbekeptin-20℃topreserve,Avoidrepeatedfreeze-thawcycles.2.Can’tdetectthesamplewhichcontainNaN3,becauseNaN3inhibitsHRPactive.Assayprocedure1.Diluteandaddsample:DiluteOriginaldensityStandardasfollowtable:0.8Eu/L5Standard150μlOriginaldensityStandard+150μlStandarddiluent0.4Eu/L4Standard150μl5Standard+150μlStandarddiluent0.2Eu/L3Standard150μl4Standard+150μlStandarddiluent0.1Eu/L2Standard150μl3Standard+150μlStandarddiluent0.05Eu/L1Standard150μl2Standard+150μlStandarddiluent2.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.RatLPS試劑盒3StepsdescriptionStandard,SamplediluentAddStandard,Samplediluent,incubatefor30minat37℃.Wash5time,AddHRP-Conjugatereagent,incubatefor30minat37℃.Wash5times,AddChromogenSolutionAandB,incubatefor30minat37℃.AddStoppSolutionReadabsorbanceat450nmwithin15mincalculateCalculateTakethestandarddensityasthehorizontal,theODvalueforthevertical,drawthestandardcurveongraphpaper,FindoutthecorrespondingdensityaccordingtothesampleODvaluebytheSamplecurve,multipliedbythedilutionmultiple,orcalculatethestraightlineregressionequationofthestandardcurvewiththestandarddensityandtheODvalue,withthe4sampleODvalueintheequation,calculatethesampledensity,multipliedbythedilutionfactor,theresultisthesampleactualdensity.Importantnotes1.Thekittakesoutfromtherefrigerationenvironment首ldbebalanced15-30minutesintheroomtemperature,ELISAplatescoatedifhasnotuseupafteropened,theplate首ldbestoredinSealedbag.2.washingbufferwillCrystallizationseparation,itcanbeheatedthewaterhelpsdissolvewhendilute.Washingdoesnotaffecttheresult.3.addSamplewithsamplerEachstep,Andproofreaditsaccuracyfrequently,avoidstheexperimentalerror.addsamplewithin5min,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.RatLPS試劑盒Storageandvalidity1.Storage:2-8℃.2.validity:sixmonths[詳細(xì)]
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2018-10-31 10:00
產(chǎn)品樣冊
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全自動電位滴定法測定疫苗佐劑中的碘值
- 2.1、儀器
JH-T6全自動電位滴定儀,鉑復(fù)合電極,分析天平等。
2.2、試劑
三氯甲烷,韋氏試劑,碘化鉀,硫代硫酸鈉標(biāo)準(zhǔn)滴定液(0.1mol/L)。
[詳細(xì)]
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2025-12-10 11:43
應(yīng)用文章
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Rat Aβ1-42 ELISA kit
- www.biokanu.comRatamyloidbetapeptide1-42(Aβ1-42)ELISAKitProd.No.3R285FROM:RBForthequantitativeinvitrodeterminationofAβ1-42concentrationsinRatsupernates,serum,plasmaandtissue.FORRESEARCHUSEONLY.NOTFORUSEINDIAGNOSTICPROCEDURES.TABLEOFCONTENTSContentsPageTABLEOFCONTENTS..2INTENDEDUSE..3PRINCIPLE..3WARNINGSANDPRECAUTIONS..4MATERIALSPROVIDEDWITHTHEKIT.7MATERIALSREQUIREDBUTNOTPROVIDED..7STORAGECONDITIONS..8REAGENTPREPARATION..9SPECIMENCOLLECTIONANDPREPARATION..9ASSAYPROCEDURE..10CALCULATIONOFRESULTS..13REFERENCES..14INTENDEDUSEAnenzymeimmunoassayforthequantitativeinvitrodiagnosticmeasurementofRatAβ1-42incellculturesupernates,serum,plasmaandtissue.PRINCIPLEThekitassayRatAβ1-42levelinthesample,usePurifiedRatAβ1-42antibodytocoatmicrotiterplatewells,makesolid-phaseantibody,thenaddAβ1-42towells,CombinedAβ1-42antibodywhichWithHRPlabeled,becomeantibody-antigen-enzyme-antibodycomplex,afterwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolorAtHRPenzyme-catalyzed,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofAβ1-42inthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.WARNINGSANDPRECAUTIONSlThiskitisforinvitrodiagnosticuseonly.Forprofessionaluseonly.lAllreagentsofthistestkitwhichcontainhumanserumorplasmahavebeentestedandconfirmednegativeforHIVI/II,HBsAgandHCVbyFDAapprovedprocedures.Allreagents,however,首ldbetreatedaspotentialbiohazardsinuseandfordisposal.lBeforestartingtheassay,readtheinstructionscompletelyandcarefully.Usethevalidversionofthepackageinsertprovidedwiththekit.Besurethateverythingisunderstood.lThemicroplatecontainssnap-offstrips.Unusedwellsmustbestoredat2°Cto8°Cinthesealedfoilpouchandusedintheframeprovided.lPipettingofsamplesandreagentsmustbedoneasquicklyaspossibleandinthesamesequenceforeachstep.lUsereservoirsonlyforsinglereagents.Thisespeciallyappliestothesubstratereservoirs.Usingareservoirfordispensingasubstratesolutionthathadpreviouslybeenusedfortheconjugatesolutionmayturnsolutioncolored.Donotpourreagentsbackintovialsasreagentcontaminationmayoccur.lMixthecontentsofthemicroplatewellsthoroughlytoensuregoodtestresults.Donotreusemicrowells.lDonotletwellsdryduringassay;addreagentsimmediatelyaftercompletingtherinsingsteps.lAllowthereagentstoreachroomtemperature(21-26°C)beforestartingthetest.Temperaturewillaffecttheabsorbancereadingsoftheassay.However,valuesforthepatientsampleswillnotbeaffected.lNeverpipetbymouthandavoidcontactofreagentsandspecimenswithskinandmucousmembranes.lDonotsmoke,eat,drinkorapplycosmeticsinareaswherespecimensorkitreagentsarehandled.lWeardisposablelatexgloveswhenhand領(lǐng)specimensandreagents.Microbialcontaminationofreagentsorspecimensmaygivefalseresults.lHand領(lǐng)首ldbedoneinaccordancewiththeproceduresdefinedbyanappropriatenationalbiohazardsafetyguidelineorregulation.lDonotusereagentsbeyondexpirydateasshownonthekitlabels.lAllindicatedvolumeshavetobeperformedaccordingtotheprotocol.Optimaltestresultsareonlyobtainedwhenusingcalibratedpipettesandmicrotiterplatereaders.lDonotmixorusecomponentsfromkitswithdifferentlotnumbers.Itisadvisednottoexchangewellsofdifferentplatesevenofthesamelot.Thekitsmayhavebeenshippedorstoredunderdifferentconditionsandthebindingcharacteristicsoftheplatesmayresultslightlydifferent.lAvoidcontactwithStopSolutioncontaining0.5MH2SO4.Itmaycauseskinirritationandburns.lSomereagentscontainProclin,BNDand/orMITaspreservatives.Incaseofcontactwitheyesorskin,flushimmediatelywithwater.lTMBsubstratehasanirritanteffectonskinandmucosa.Incaseofpossiblecontact,washeyeswithanabundantvolumeofwaterandskinwithsoapandabundantwater.Washcontaminatedobjectsbeforereusingthem.Ifinhaled,takethepersontoopenair.lChemicalsandpreparedorusedreagentshavetobetreatedashazardouswasteaccordingtothenationalbiohazardsafetyguidelineorregulation.lForinformationonhazardoussubstancesincludedinthekitpleaserefertoMaterialSafetyDataSheetsMATERIALSPROVIDEDWITHTHEKITMaterialsprovidedwiththekit96determinationsStorageUsermanual1Closureplatemembrane2Sealedbags1Microelisastripplate12-8℃Standard:900pg/ml0.5ml×1bottle2-8℃Standarddiluent1.5ml×1bottle2-8℃HRP-Conjugatereagent6ml×1bottle2-8℃Samplediluent6ml×1bottle2-8℃ChromogenSolutionA6ml×1bottle2-8℃ChromogenSolutionB6ml×1bottle2-8℃StopSolution6ml×1bottle2-8℃washsolution30×20ml×1bottle2-8℃MATERIALSREQUIREDBUTNOTPROVIDEDlMicroplatereadercapableofmeasuringabsorbanceat450nm.lPrecisionpipettestodeliver2mlto1mlvolumes.l100mland1litergraduatedcylinders.lCalibratedadjustableprecisionpipettes,preferablywithdisposableplastictips.(Amanifoldmulti-channelpipetteisdesirableforlargeassays.)lAbsorbentpaper.l37°Cincubator.lDistilledordeionizedwater.lDataanalysisandgraphingsoftware.Graphpaper:linear(Cartesian),log-logorsemi-log,orlog-logitasdesired.lTubestopreparestandardorsampledilutions.STORAGECONDITIONSuWhenstoredat2°Cto8°Cunopenedreagentswillretainreactivityuntilexpirationdate.uDonotusereagentsbeyondthisdate.Openedreagentsmustbestoredat2°Cto8°C.uMicrotiterwellsmustbestoredat2°Cto8°C.Oncethefoilbaghasbeenopened,care首ldbetakentocloseittightlyagain.uOpenedkitsretainactivityfor8weeksifstoredasdescribedabove.REAGENTPREPARATIONBringallreagentstoroomtemperaturebeforeuseSPECIMENCOLLECTIONANDPREPARATIONSerum-Useaserumseparatortube(SST)andallowsamplestoclotfor30minutesbeforecentrifugationfor15minutesatapproximately1000xg.Removeserumandassayimmediatelyoraliquotandstoresamplesat-20°Cor-80°C.Plasma-CollectplasmausingEDTAorheparinasananticoagulant.Centrifugesamplesfor15minutesat1000xgat2-8°Cwithin30minutesofcollection.Storesamplesat-20°Cor-80°C.Avoidrepeatedfreeze-thawcycles.Cellculturefluidandotherbiologicalfluids-Removeparticulatesbycentrifugationandassayimmediatelyoraliquotandstoresamplesat-20°Cor-80°C.Avoidrepeatedfreeze-thawcyclesASSAYPROCEDUREuGeneralRemarkslAllreagentsandspecimensmustbeallowedtocometoroomtemperaturebeforeuse.Allreagentsmustbemixedwithoutfoaming.lOncethetesthasbeenstarted,allsteps首ldbecompletedwithoutinterruption.lUsenewdisposalplasticpipettetipsforeachstandard,controlorsampleinordertoavoidcrosscontamination.lAbsorbanceisafunctionoftheincubationtimeandtemperature.Beforestartingtheassay,itisrecommendedthatallreagentsareready,capsremoved,allneededwellssecuredinholder,etc.Thiswillensureequalelapsedtimeforeachpipettingstepwithoutinterruption.lAsageneralruletheenzymaticreactionislinearlyproportionaltotimeandtemperature.lDetermineabsorptionwithanELISAreaderat450nmagainst620nmasreference.Ifnoreferencewavelengthisavailable,readonlyat450nm.Iftheextinctionofthehigheststandardexceedsthemeasurementrangeofthephotometer,absorptionmustbemeasuredimmediatelyat405nmagainst620nmasreference.uAssayProcedure1.DiluteandaddsampletoStandard:set10StandardwellsontheELISAplatescoated,addStandard100μltothefirstandthesecondwell,thenaddStandarddilution50μltothefirstandthesecondwell,mix;takeout100μlformthefirstandthesecondwellthenaddittothethirdandtheforthwellseparately.thenaddStandarddilution50μltothethirdandtheforthwell,mix;thentakeout50μlfromthethirdandtheforthwelldiscard,add50μltothefifthandthesixthwell,thenaddStandarddilution50μltothefifthandthesixthwell,mix;takeout50μlfromthefifthandthesixthwellandaddtotheseventhandtheeighthwell,thenaddStandarddilution50μltotheseventhandtheeighthwell,mix;takeout50μlfromtheseventhandtheeighthwellandaddtotheninthandthetenthwell,addStandarddilution50μltotheninthandthetenthwell,mix,takeout50μlfromtheninthandthetenthwelldiscard(addSample50μltoeachwellafterDiluting,(density:600pg/ml,400pg/ml,200pg/ml,100pg/ml,50pg/ml).50pg/ml100pg/ml600pg/ml200pg/ml900pg/ml400pg/ml2.addsample:Setblankwellsseparately(blankcomparisonwellsdon’taddsampleandHRP-Conjugatereagent,othereachstepoperationissame).testingsamplewell.addSampledilution40μltotestingsamplewell,thenaddtestingsample10μl(samplefinaldilutionis5-fold),addsampletowells,don’ttouchthewellwallasfaraspossible,andGentlymix.3.Incubate:AfterclosingplatewithClosureplatemembrane,incubatefor30minat37℃.4.Configurateliquid:30-foldwashsolutiondiluted30-foldwithdistilledwaterandreserve.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.CALCULATIONOFRESULTSlCalculatetheaverageabsorbancevaluesforeachsetofstandards,controlsandpatientsamples.lConstructastandardcurvebyplottingthemeanabsorbanceobtainedfromeachstandardagainstits.lconcentrationwithabsorbancevalueonthevertical(Y)axisandconcentrationonthehorizontal(X)axis.lUsingthemeanabsorbancevalueforeachsampledeterminethecorrespondingconcentrationfromthestandardcurve.lAutomatedmethod:TheresultsintheIFUhavebeencalculatedautomaticallyusinga4PL.l(4ParameterLogistics)curvefit.4ParameterLogisticsisthepreferredcalculationmethod.Otherdata.lreductionfunctionsmaygiveslightlydifferentresults.lTheconcentrationofthesamplescanbereaddirectlyfromthisstandardcurve.Sampleswith.lconcentrationshigherthanthatofthehigheststandardhavetobefurtherdiluted.Forthecalculationof.ltheconcentrationsthisdilutionfactorhastobetakenintoaccount.REFERENCESREF:Cat.-No.:/Kat.-Nr.:/No.-Cat.:/Cat.-No.:/N.Cat.:/N.CatLOT:Lot-No.:/Chargen-Bez.:/No.Lot:/Lot-No.:/LoteN.:/Lotton.::No.ofTests:/Kitgre:/Nb.deTests:/No.deDeterm.:/N.deTestes:/Quantitàdeitests::Keepawayfromheatordirectsunlight./VorHitzeunddirekterSonneneinstrahlungschützen./Garderàl’abridelachaleuretdetouteexpositionlumineuse./Manténgasealejadodelcalorolaluzsolardirecta./Manterlongedocalorouluzsolardirecta./Nonesporreairaggisolari.:Readinstructionsbeforeuse./Arbeitsanleitunglesen./Lirelafichetechniqueavantemploi./Lealasinstruccionesantesdeusar./Lerasinstruesantesdeusar./Leggereleistruzioniprimadell’uso.:Storeat:/Lagernbei:/Stockerà:/Almacenea:/Armazenara:/Conservarea:[詳細(xì)]
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2018-09-22 10:00
產(chǎn)品樣冊
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甲流疫苗中的鋁佐劑粒度控制
- 甲流疫苗中的鋁佐劑粒度控制[詳細(xì)]
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2024-09-20 14:20
應(yīng)用文章
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電位滴定法測定動物疫苗佐劑中的酸值含量
- 一、前言
佐劑是疫苗的一種添加劑,是能夠非特異性地改變或增強(qiáng)機(jī)體對抗原的特異性免疫應(yīng)答、發(fā)揮輔助作 用的一類物質(zhì)。近年來為適應(yīng)新型疫苗的需求,佐劑已經(jīng)從傳統(tǒng)、單一的形式向新型、多元化形式發(fā)展, 理想的[詳細(xì)]
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2025-07-02 14:54
應(yīng)用文章
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干燥箱的運(yùn)用流程
- 干燥箱的運(yùn)用流程[詳細(xì)]
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2013-08-01 00:00
期刊論文
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干燥箱的操作流程
- 干燥箱的操作流程[詳細(xì)]
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2024-09-15 17:51
應(yīng)用文章
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全自動電位滴定法測定疫苗佐劑羥值
- 2.1、儀器
JH-T6 全自動電位滴定儀,PH 復(fù)合電極,水浴鍋,分析天平等。
2.2、試劑
?;噭ㄈ妆交撬?14.4g, 置干燥的 500ml 具塞錐形瓶中,加醋酸乙酯 360ml,[詳細(xì)]
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2025-12-08 10:29
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