TheThermoScientificPierceBCAProteinAssayKitisatwo-component,high-precision,detergent-compatIBLeassayreagentsettomeasure(A562nm)totalproteinconcentrationcomparedtoaproteinstandard.
FeaturesoftheBCAProteinAssayKit:
• Colorimetric—estimatevisuallyormeasurewithastandardspectrophotometerorplatereader(562nm)
• Excellentuniformity—exhibitslessprotein-to-proteinvariationthandye-bindingmethods
• Compatible—unaffectedbytypicalconcentrationsofmostionicandnonionicdetergents
• Moderatelyfast—mucheasierandfourtimesfasterthantheclassicalLowrymethod
• Highlinearity—linearworkingrangeforBSAequals20to2000µg/mL
• Sensitive—detectdownto5µg/mLwiththeenhancedprotocol
Usedinmorelabsthananyotherdetergent-compatibleproteinassay,PierceBCAReagentsprovideaccuratedeterminationofproteinconcentrationwithmostsampletypesencounteredinproteinresearch.ThePierceBCAAssaycanbeusedtoassessyieldsinwholecelllysatesandaffinity-columnfractions,aswellastomonitorproteincontaminationinindustrialapplications.Comparedtomostdye-bindingmethods,theBCAAssayisaffectedmuchlessbyproteincompositionaldifferences,providinggreaterprotein-to-proteinuniformity.
Applications:
•Studyingprotein:proteininteractions
•Measuringcolumnfractionsafteraffinitychromatography
•Estimatingpercentrecoveryofmembraneproteinsfromcellextracts
•High-throughputscreeningoffusionproteins
HowtheBCAProteinAssayDetectsProtein:
TheBCAProteinAssaycombinesthewell-knownreductionofCu2+ toCu1+ byproteininanalkalinemediumwiththehighlysensitiveandselectivecolorimetricdetectionofthecuprouscation(Cu1+)bybicinchoninicacid.Thefirststepisthechelationofcopperwithproteininanalkalineenvironmenttoformalightbluecomplex.Inthisreaction,knownasthebiuretreaction,peptidescontainingthreeormoreaminoacidresiduesformacoloredchelatecomplexwithcupricionsinanalkalineenvironmentcontainingsodiumpotassiumtartrate.
Inthesecondstepofthecolordevelopmentreaction,bicinchoninicacid(BCA)reactswiththereduced(cuprous)cationthatwasformedinstepone.Theintensepurple-coloredreactionproductresultsfromthechelationoftwomoleculesofBCAwithonecuprousion.TheBCA/coppercomplexiswater-solubleandexhibitsastronglinearabsorbanceat562nmwithincreasingproteinconcentrations.TheBCAreagentisapproximately100timesmoresensitive(lowerlimitofdetection)thanthepalebluecolorofthefirstreaction.
ThereactionthatleadstoBCAcolorformationisstronglyinfluencedbyfouraminoacidresidues(cysteineorcystine,tyrosine,andtryptophan)intheaminoacidsequenceoftheprotein.However,unliketheCoomassiedye-bindingmethods,theuniversalpeptidebackbonealsocontributestocolorformation,helpingtominimizevariABIlitycausedbyproteincompositionaldifferences.
Pierce属于世界著名Fisher Biosciences公司旗下的Pierce生物技术公司最近宣布与GenTel BioSurfaces公司达成协议:代理销售其公司的GenTel PATH品牌的超薄硝酸纤维膜玻片,并且获得利用GenTel技术授权,可以利用此技术开发新的应用于生命科学研究和药物开发的蛋白芯片。
GenTel玻片是专门为蛋白芯片而设计的,它采用超薄的硝酸纤维膜放置在一个标准的3" x 1"玻璃片上的设计。这种硝酸纤维膜比传统的要薄上50倍,可以大大改善实验的结果。GenTel技术玻片可以用于许多用途。
采用GenTel技术玻片拥有较高的结合能力,并且较低的背景,这样可以使之后的荧光检测变得更加灵敏。另外,这类玻片和Pierce的化学发光检测系统也是可以兼容的,这使更高灵敏度的检测成为可以,满足用户更挑剔的需求。