WesternBlottingorimmunoblotting(WesternblottingwasdubbedbecauseitissimilartoSouthernblottingwhichwasinventedbyandnamedaftertheinventorE.M.Southern)allowsinvestigatorstodetermine,withaspecificprimaryantibody,therelativeamountsoftheproteinpresentindifferentsamples.Briefly,1)Samplesarepreparedfromtissuesorcellsthatarehomogenizedinabufferthatprotectstheproteinofinterestfromdegradation;2)ThesampleisseparatedusingSDS-PAGEandthentransferredtoamembranefordetection;3)Themembraneisincubatedwithagenericprotein(suchasmilkproteins)tobindtoanyremainingstickyplacesonthemembrane.Aprimaryantibodyisthenaddedtothesolutionwhichisabletobindtoitsspecificprotein;4)Asecondaryantibody-enzymeconjugate,whichrecognizestheprimaryantibodyisaddedtofindlocationswheretheprimaryantibodybound.

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