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  • Quantitation of protein on gels and blots by infrared fluorescence of Coomassie blue and Fast Green.

Quantitation of protein on gels and blots by infrared fluorescence of Coomassie blue and Fast Green.

Analytical biochemistry (2005-12-13)
Shen Luo, Nancy B Wehr, Rodney L Levine
ABSTRACT

Coomassie blue staining of gels and blots is commonly employed for detection and quantitation of proteins by densitometry. We found that Coomassie blue or Fast Green FCF bound to protein fluoresces in the near infrared. We took advantage of this property to develop a rapid and sensitive method for detection and quantitation of proteins in gels and on blots. The fluorescence response is quantitative for protein content between 10 ng and 20 microg per band or spot. Staining and destaining require only 30 min, and the method is compatible with subsequent immunodetection.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Brilliant Blue R, 250, suitable for microscopy
Sigma-Aldrich
Brilliant Blue R Concentrate, suitable for SDS-PAGE, methanol solution
Sigma-Aldrich
Brilliant Blue R, Dye content ~50 %, Technical grade
Sigma-Aldrich
Brilliant Blue R Staining Solution, suitable for (for immunoelectrophoresis protein staining)
Sigma-Aldrich
Brilliant Blue R, pure
Supelco
Fast Green FCF, analytical standard
Sigma-Aldrich
Fast Green FCF, Dye content ≥85 %
Sigma-Aldrich
Fast Green FCF, certified by the BSC