Introduction

Proteomicsisafieldofstudythatfocusesonthepropertiesofproteinsnamely
theirstructure,location,functionandinteractionwithotherbiomolecules(BioRad,n.d.).
Theprerequisitetoperformingsuchstudiesisproteinpurificationandthesuccessof
whichisverifiedbyproteinquantitation.
Protein quantitation is a crucial step before processing protein samples for
isolation, separation and analysis by chromatography, electrophoresis and
immunochemicaltechniques(Hayworth,n.d.).Determiningproteinconcentrationisan
integralpartofworkflowsinbiochemistryresearchlabsforvariousapplicationsranging
from enzymatic activities to releasing data for the formulation of biopharmaceutical
drugs (Noble & Bailey, 2009). Different assays for protein quantitation are used
depending on the amount of purified protein available and the preferred accuracy.
Unfortunately,noparticularreagentorassayisproteinspecificandnoneisuniformly
sensitivetoallproteins(Hayworth,n.d.).However,whenthereisaneedtomeasurethe
total protein concentration, the assays recommended are those compatible with most
proteinsamples;oneofwhichistheCoomassie(Bradford)Assay(Hayworth,n.d.).
TheconceptbehindtheBradfordassayliesonthecolordevelopmentformed
whenthedye,CoomassieBlueG250,bindstospecificRgroupsorsidechainsofthe
amino acids (BioRad, n.d.). Such technique is equipped with the technicalities of
biophotonics or the study of biomolecules with photon detection. Upon binding, the
samplesolutionturnsblue.Theintensityofthebluecolorhasapositivecorrelationwith
proteinconcentration,whichrendersqualitativeestimationpossiblewiththisassay,if
standardsaregiven.Thebestestimateofproteincontentcanbeobtainedbymeasuring
theabsorbancevaluesoftheproteinsamplesusingaspectrophotometer(BioRad,n.d.).
Intheexperiment,Bradfordassaywasemployedtodeterminetheproteincontent
ofeggandmilksamples.Thisexperimentaimedtogenerateastandardcurveandfrom
this,theequationforobtainingproteinconcentrationwasderived.Lastly,thisactivity
aimedtodemonstratetheconceptofbiophotonicsandtoexplaintheeffectofreagents
andotherfactorsontheefficacyoftheBradfordassayinproteinquantitation.
References:
BioRad.(n.d.).GotProtein?KitStudentManual.RetrievedNovember29,2014,from
http://www.biorad.com/webroot/web/pdf/lse/literature/10006836A.pdf
Hayworth,D.(n.d.).OverviewofProteinAssaysMethods.RetrievedNovember29,
2014,fromhttp://www.piercenet.com/method/overviewproteinassays

Noble,J.E.,&Bailey,M.J.A.(2009).Chapter8:QuantitationofProtein.Methodsin
Enzymology,463,7395.AcademicPress.RetrievedNovember29,2014,from
http://www.sciencedirect.com/science/article/pii/S0076687909630081