BCA蛋白濃度測(cè)定試劑盒(增強(qiáng)型)

BCA蛋白濃度測(cè)定試劑盒(增強(qiáng)型)BCA蛋白濃度測(cè)定試劑盒(增強(qiáng)型)BCA蛋白濃度測(cè)定試劑盒(增強(qiáng)型)產(chǎn)品編號(hào)產(chǎn)品名稱包裝蛋白濃度測(cè)定試劑盒(增強(qiáng)型次產(chǎn)品簡(jiǎn)介：BCA蛋白濃度測(cè)定試劑盒(增強(qiáng)型)(EnhancedBCAProteinAssayKit)是根據(jù)目前世界上最常用的兩種蛋白濃度檢測(cè)方法之一BCA法研制而成，實(shí)現(xiàn)了蛋白濃度測(cè)定的簡(jiǎn)單，高穩(wěn)定性，高靈敏度和高兼容性。和碧云天生產(chǎn)的普通BCA蛋白濃度測(cè)定試劑盒相比，靈敏度更高，檢測(cè)濃度下限達(dá)到10g/ml，最小檢測(cè)蛋白量達(dá)到0.2g，待測(cè)樣品體積為1-20l。和碧云天生產(chǎn)的普通BCA蛋白濃度測(cè)定試劑盒相比，顯色速度更快，相同的樣品孵育較短時(shí)間即可進(jìn)行吸光度測(cè)定。在20-1000g/ml濃度范圍內(nèi)有較好的線性關(guān)系。BCA法測(cè)定蛋白濃度不受絕大部分樣品中的化學(xué)物質(zhì)的影響，可以兼容樣品中高達(dá)5%的SDS，5%的TritonX-100，5%的Tween20,60,80。但受螯合劑和略高濃度的還原劑的影響，需確保EDTA低于10mM，無EGTA，二硫蘇糖醇低于1mM，β-巰基乙醇低于0.01%。不適用BCA法時(shí)建議試用碧云天生產(chǎn)的Bradford蛋白濃度測(cè)定試劑盒(P0006)。BCA蛋白濃度測(cè)定試劑盒(增強(qiáng)型)對(duì)樣品中各種物質(zhì)的詳細(xì)的兼容性和普通的BCA蛋白濃度測(cè)定試劑盒相同，請(qǐng)參考碧云天如下網(wǎng)頁:/CompatibilityChartForBCAKit.pdf每個(gè)試劑盒可以檢測(cè)5000個(gè)樣品。包裝清單：產(chǎn)品編號(hào)產(chǎn)品名稱包裝試劑A500ml/瓶，共2瓶P0009-2BCA試劑B30mlP0009-3蛋白標(biāo)準(zhǔn)管，共2管P0009-4蛋白標(biāo)準(zhǔn)配制液5ml―說明書1份保存條件：室溫保存。蛋白標(biāo)準(zhǔn)配制成溶液后-20℃凍存。注意事項(xiàng)：需酶標(biāo)儀一臺(tái)，測(cè)定波長為540-595nm之間，562nm最佳。需96孔板。如果沒有酶標(biāo)儀，也可以使用普通的分光光度計(jì)測(cè)定，但測(cè)定時(shí)，需根據(jù)比色皿的最小檢測(cè)體積，適當(dāng)加大BCA工作液的用量使不小于最小檢測(cè)體積，樣品和標(biāo)準(zhǔn)品的用量可相應(yīng)按比例放大也可不變。使用分光光度計(jì)測(cè)定蛋白濃度時(shí)，每個(gè)試劑盒可以測(cè)定的樣品數(shù)量可能會(huì)顯著減少。如發(fā)現(xiàn)樣品稀釋液或裂解液本身就有較高背景，請(qǐng)?jiān)囉帽淘铺焐a(chǎn)的Bradford蛋白濃度測(cè)定試劑盒(P0006)。為了加快BCA法測(cè)定蛋白濃度的速度可以適當(dāng)用微波爐加熱，但是切勿過熱。EDTA濃度必須小于10mM，不兼容EGTA。不適用BCA法時(shí)，請(qǐng)?jiān)囉帽淘铺焐a(chǎn)的Bradford蛋白濃度測(cè)定試劑盒(P0006)。為了您的安全和健康，請(qǐng)穿實(shí)驗(yàn)服并戴一次性手套操作。使用說明：1.取1.2ml蛋白標(biāo)準(zhǔn)配制液加入到一管蛋白標(biāo)準(zhǔn)(30mgBSA)中，充分溶解后配制成25mg/ml的蛋白標(biāo)準(zhǔn)溶液。配制后可立即使用，也可以-20℃長期保存。2.取適量25mg/ml蛋白標(biāo)準(zhǔn)，稀釋至終濃度為0.5mg/ml。例如取20l25mg/ml蛋白標(biāo)準(zhǔn)，加入980l稀釋液即可配制成0.5mg/ml蛋白標(biāo)準(zhǔn)。蛋白樣品在什么溶液中，標(biāo)準(zhǔn)品也宜用什么溶液稀釋。但是為了簡(jiǎn)便起見，也可以用0.9%NaCl或PBS稀釋標(biāo)準(zhǔn)品。稀釋后的0.5mg/ml蛋白標(biāo)準(zhǔn)也可以-20℃長期保存。3.根據(jù)樣品數(shù)量，按50體積BCA試劑A加1體積BCA試劑B(50:1)配制適量BCA工作液，充分混勻。例如5mlBCA試劑A加100lBCA試劑B，混勻，配制成5.1mlBCA工作液。BCA工作液室溫24小時(shí)內(nèi)穩(wěn)定。4.將標(biāo)準(zhǔn)品按0,1,2,4,8,12,16,20l加到96孔板的標(biāo)準(zhǔn)品孔中，加標(biāo)準(zhǔn)品稀釋液補(bǔ)足到20l。5.加適當(dāng)體積樣品到96孔板的樣品孔中，加標(biāo)準(zhǔn)品稀釋液到20l。6.各孔加入200lBCA工作液，37℃放置20-30分鐘。注:也可以室溫放置2小時(shí)，或60℃放置30分鐘。BCA法測(cè)定蛋白濃度時(shí)，顏色會(huì)隨著時(shí)間的延長不斷加深。并且顯色反BCA蛋白濃度測(cè)定試劑盒(增強(qiáng)型)應(yīng)會(huì)因溫度升高而加快。如果濃度較低，適合在較高溫度孵育，或適當(dāng)延長孵育時(shí)間。7.測(cè)定A562，540-595nm之間的波長也可接受。根據(jù)標(biāo)準(zhǔn)曲線計(jì)算出樣品的蛋白濃度。常見問題：1.測(cè)定標(biāo)準(zhǔn)曲線時(shí)發(fā)現(xiàn)隨著標(biāo)準(zhǔn)品濃度的增加吸光度或顏色沒有明顯變化?？赡艿脑蚴菢悠分泻袊?yán)重干擾BCA法測(cè)定蛋白濃度的物質(zhì)，詳細(xì)的BCA法的兼容性列表請(qǐng)參考碧云天如下網(wǎng)頁:/CompatibilityChartForBCAKit.pdf2.是否每次測(cè)定時(shí)都需要做標(biāo)準(zhǔn)曲線？建議每次測(cè)定時(shí)都做標(biāo)準(zhǔn)曲線。因?yàn)锽CA法測(cè)定時(shí)顏色會(huì)隨著時(shí)間的延長不斷加深，并且顯色反應(yīng)的速度和溫度有關(guān)，所以除非精確控制顯色反應(yīng)的時(shí)間和溫度，否則如需精確測(cè)定宜每次都做標(biāo)準(zhǔn)曲線。使用本產(chǎn)品的文獻(xiàn)：1.LiuMJ,WangZ,LiHX,WuRC,LiuYZ,WuQY.MitochondrialdysfunctionasanearlyeventintheprocessofapoptosisinducedbywoodfordinIinhumanleukemiaK562cells.ToxicolApplPharmacol.2022年Jan15;194(2):141-55.2.WuRC,ChenDF,LiuMJ,WangZ.DualeffectsofcycloheximideonU937apoptosisinducedbyitscombinationwithVP-16.BiolPharmBull.2022年Jul;27(7):1075-80.3.ZhengCH,GaoJQ,ZhangYP,LiangWQ.Aproteindeliverysystem:biodegradablealginate-chitosan-poly(lactic-co-glycolicacid)compositemicrospheres.BiochemBiophysResCommun.2022年Oct29;323(4):1321-7.4.R.-C.Wu,Z.Wang,M.-J.Liu,D.-F.ChenandX.-S.Yue.β2-integrinsmediateanovelformofchemoresistanceincycloheximide-inducedU937apoptosis.Cell.Mol.LifeSci.2022年,61(16):2071C2082.5.劉軍丁勁薛采芳李英輝宮衛(wèi)東趙亞黃豫曉。帶有蛋白轉(zhuǎn)導(dǎo)域的靶向核糖核酸酶對(duì)乙型肝炎病毒復(fù)制的影響。中華肝臟病雜志2022年第6期第377頁。6.Zhong-shuLiang,KanYangandBen-meiChen.CholesterolinU937foamcellsassayedinliquidchromatographic-massspectrometry.ChinJModMed;2022年,14(20):51-56.7.LiuMJ,YuePY,WangZ,WongRN.MethylprotodioscininducesG2/MarrestandapoptosisinK562cellswiththehyperpolarizationofmitochondria.CancerLett.2022年,224:229C241.8.LiuMJ,WangZ,JuY,WongRN,WuQY.DiosgenininducescellcyclearrestandapoptosisinhumanleukemiaK562cellswiththedisruptionofCa2+homeostasis.CancerChemotherPharmacol.2022年Jan;55(1):79-90.9.JinDing,JunLiu,Cai-FangXue,Ying-HuiLi,YaZhao,JunChen,Yu-XiaoHuang,Zhong-XiangLiu.TatPTDcanintroduceHBVtargetedribonucleaseintohepatocytes.WorldChinJDigestol.2022年april15;13(8):958-962.10.ZhaoY,GuanH,LiuSF,WuRC,WangZ.OverexpressionofQMinducescelldifferentiationandmineralizationinMC3T3-E1.BiolPharmBull.2022年Aug;28(8):1371-6.11.ZhangLF,PengSQ,WangS.Influenceoflead(Pb2+)onthereactionsofinvitroculturedrataortato5-hydroxytryptamine.ToxicolLett.2022年Oct15;159(1):71-82.12.Le-FengZhang,Shuang-QingPeng,ShengWang.Influenceoflead(Pb2+)onthereactionsofinvitroculturedrataortato5-hydroxytryptamine.ToxicologyLetters159(2022年)71C82.13.GaoJianmei,WanYanzhen,HanJun,WangXiaofan,ChenLan,NieKai,ZhouWei,DongXiaoping.InfluenceoftheNumbersofOctapeptideRepeatswithinN2terminusofRecombinantHumanPrPProteinsontheProteaseResi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