Quantitation of protein on gels and blots by infrared fluorescence of Coomassie blue and Fast Green.
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.
Luo
Shen
S
Laboratory of Biochemistry, National Heart, Lung, and Blood Institute, Bethesda, MD 20892, USA.
Wehr
Nancy B
NB
Levine
Rodney L
RL
D000180:Acrylic Resins / Q000737:chemistry; D000818:Animals; D005782:Gels / Q000737:chemistry*; D005974:Glutamate-Ammonia Ligase / Q000032:analysis; D007259:Infrared Rays; D008086:Lissamine Green Dyes / Q000737:chemistry*; D008567:Membranes, Artificial; D051150:Methionine Sulfoxide Reductases; D051379:Mice; D009113:Muramidase / Q000032:analysis; D010088:Oxidoreductases / Q000032:analysis; D011145:Polyvinyls; D011506:Proteins / Q000032:analysis*; D012394:Rosaniline Dyes / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000032:analysis; D013050:Spectrometry, Fluorescence / Q000379:methods*; D005465:Fluorocarbon Polymers
D000180:Acrylic Resins; D005782:Gels; D005974:Glutamate-Ammonia Ligase; D008086:Lissamine Green Dyes; D008567:Membranes, Artificial; D051150:Methionine Sulfoxide Reductases; D009113:Muramidase; D010088:Oxidoreductases; D011145:Polyvinyls; D011506:Proteins; D012394:Rosaniline Dyes; D012710:Serum Albumin, Bovine; C016680:polyacrylamide gels; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers; C007704:Fast Green FCF; C024757:Coomassie brilliant blue R; C027219:methionine sulfoxide reductase