描述
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.
Formoreinformation,seethearticle"ChemistryofProteinAssays"intheProteinMethodsLibrary.
RelatedProducts
Pierce?BCAProteinAssayReagentA
Pierce?BCAProteinAssayReagentB
Pierce?BCASolid
AmpuleBreakers
規(guī)格
AssayType: | ProteinQuantitationAssay |
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DetectionMethod: | Colorimetric |
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ForUseWith(Equipment): | MicroplateReader, Spectrophotometer |
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ProductLine: | Pierce? |
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ProductSize: | 1L |
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Technique: | Absorbance, Solution-basedDetection |
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內(nèi)容及儲(chǔ)存
SufficientFor:500tubeassaysor5000microplateassays
?BCAReagentA,2x500mL
?BCAReagentB,25mL
?AlbuminStandardAmpules,2mg/mL,10x1mL
Storeatroomtemperature