Alternative Method for Quantifying Protein Expression in Western Blotting Analysis

Abstract: Image densitometry has been widely used to quantify specific cellular protein expression in Western blotting analysis， but still there exists an unresolved problem of sensitivity. Furthermore， the use of expensive reagents and complex equipment limits its application. Here， we establish an improved method for quantitative examination by using alkaline phosphatase-labeled secondary antibody in combination with substrate compound naphthol AS-MX phosphate and chromagen 4-chloro-2-methylbenzenediazonium salt (fast red TR). The final colorization on blotted membrane is eluted using an organic solvent dimethylsulfoxide (DMSO) and quantified by reading absorptance at 492 nm. Our results prove that this technique is more simplified， fast and economic， without loss of its sensitivity.

CSTR: 32200.14.cjcb.2007.02.0025