Highly increased detection of silver stained protein bands in polyacrylamide gels with thermo-optical methods

Abstract

Sodium dodecyl sulfate polyacrylamide gel electrophoresis is a well-known technique to separate proteins by their molecular weight. After electrophoresis, the gels are commonly stained for protein band analysis with silver stain; this allows the detection of protein loads to about 1 ng. To increase the detection sensitivity of the protein bands down in the subnanogram level, a sensor has been developed based on the thermal lens effect to scan and quantify protein loads which would remain undetected using the standard imaging systems. The thermal lens sensor is equipped with a 450 nm diode pump laser modulated at 1 Hz and a HeNe probe laser mounted in collinear geometry. The sensor could detect protein bands of 0.05 ng when the gel was soaked in methanol/water and 0.1 ng in water. The limit of detection ranged from 8 to 20 pg, depending on the soaking medium and the staining efficiency. Thus, the detection of silver stain by thermal lens effect results 10 to 20 times more sensitive than the standard colorimetric method.