CBB staining protocol with higher sensitivity and mass spectrometric compatibility.
Various CBB-based methods for staining proteins separated by 2-D gel electrophoresis were compared with regard to sensitivity and resolution. A modified Kang's CBB staining protocol, which we have modified, includes phosphoric acid in a concentration of 8% instead of the original 2%. This proved to be the best approach. Protein amounts as low as 2 ng and approximately 2300 spots in the gel can be detected by employing this protocol. The modified procedure takes less time to carry out. Moreover, this practice is more sensitive and resolves more protein spots than most protocols reported to date and is compatible with subsequent mass spectrometric analysis.
Pink
Mario
M
Institute of Hygiene and Occupational Medicine, University Hospital Essen, D-45122 Essen, Germany.
Verma
Nisha
N
Rettenmeier
Albert W
AW
Schmitz-Spanke
Simone
S
D002457:Cell Extracts / Q000737:chemistry; D045744:Cell Line, Tumor; D003102:Colloids / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D013058:Mass Spectrometry / Q000379:methods*; D010756:Phosphoric Acids / Q000737:chemistry; D020543:Proteome / Q000737:chemistry*; D015203:Reproducibility of Results; D012394:Rosaniline Dyes / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods*
D002457:Cell Extracts; D003102:Colloids; D010756:Phosphoric Acids; D020543:Proteome; D012394:Rosaniline Dyes; C030242:phosphoric acid; C004692:coomassie Brilliant Blue