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

Giulia Mazza; Thomas Posnicek; Martin Brandl

doi:10.1117/12.2244491

10 November 2016 Highly increased detection of silver stained protein bands in polyacrylamide gels with thermo-optical methods

Giulia Mazza, Thomas Posnicek, Martin Brandl

Author Affiliations +

Proceedings Volume 10161, 14th International Conference on Optical and Electronic Sensors; 101610H (2016) https://doi.org/10.1117/12.2244491
Event: 14th International Conference on Optical and Electronic Sensors, 2016, Gdansk, Poland

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.

Citation Download Citation

Giulia Mazza, Thomas Posnicek, and Martin Brandl "Highly increased detection of silver stained protein bands in polyacrylamide gels with thermo-optical methods", Proc. SPIE 10161, 14th International Conference on Optical and Electronic Sensors, 101610H (10 November 2016); https://doi.org/10.1117/12.2244491

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