Simple, Time-Saving Dye Staining of Proteins for Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis Using Coomassie Blue
نویسندگان
چکیده
A fixation-free and fast protein-staining method for sodium dodecyl sulfate-polyacrylamide gel electrophoresis using Coomassie blue is described. The protocol comprises staining and quick washing steps, which can be completed in 0.5 h. It has a sensitivity of 10 ng, comparable with that of conventional Coomassie Brilliant Blue G staining with phosphoric acid in the staining solution. In addition, the dye stain does not contain any amount of acid and methanol, such as phosphoric acid. Considering the speed, simplicity, and low cost, the dye stain may be of more practical value than other dye-based protein stains in routine proteomic research.
منابع مشابه
A convenient procedure for transfer blotting of coomassie blue stained proteins from PAGE gels to transparencies.
Proteins stained with Coomassie Brilliant Blue I were transferred effectively by blotting from polyacrylamide gel electrophoresis (PAGE) gels to transparencies of the type used in plain-paper copiers. The details of the original electropherogram were retained on transfer and did not fade over a period of three years. Both the protein and the associated dye transfer; however, protein does not tr...
متن کاملSimplified sodium dodecyl sulfate-polyacrylamide gel electrophoresis of unconcentrated urine with enhanced resolution and detection sensitivity.
We applied a simple sodium dodecyl sulfate-polyacrylamide gel electrophoresis method to urine. The method, developed for serum protein analysis (Clin Chem 1984;30:475-9), has a high sample throughput and gives excellent resolution with unconcentrated urine. It clearly distinguishes and characterizes proteinuric urine (7.5 microL) by Coomassie Blue staining and gives complex silver-stained patte...
متن کاملRapid microprocedure for isolating detergent-insoluble outer membrane proteins from Haemophilus species.
A rapid microprocedure for isolating detergent (sodium N-lauroyl sarcosinate)-insoluble major outer membrane proteins from Haemophilus species produced results qualitatively identical to those obtained with a commonly used preparative isolation procedure. Proteins isolated by both procedures were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis after staining with Coomassie...
متن کاملComparative advantages of imidazole-sodium dodecyl sulfate-zinc reverse staining in polyacrylamide gels.
Polyacrylamide gel electrophoresis (PAGE), which is widely used to assess protein purity and molecular weight [1], is best combined with a highly sensitive, reproducible, inexpensive, rapid, and easy to perform protein staining/detection method. Coomassie brilliant blue (CBB)1 staining is a popular method, known for its simplicity, economy, and compatibility with downstream analysis but relativ...
متن کاملMolecular-weight estimates of milk-fat-globule-membrane protein-sodium dodecyl sulphate complexes by electrophoresis in gradient acrylamide gels.
The molecular weights of milk-fat-globule-membrane proteins solubilized in sodium dodecyl sulphate were estimated by gradient gel electrophoresis. Standard curves were calibrated from both protein and glycoprotein markers of known molecular weight. Six major proteins were observed with Coomassie Blue staining and six with periodic acid-Schiff staining. The behaviour of the membrane proteins and...
متن کامل