Coomassie Brilliant Blue R-250 Protein Stain

1.Quantitation of proteins by elution of Coomassie brilliant blue R from stained bands after sodium dodecyl sulfate-polyacrylamide gel electrophoresis/PMID 2424336; Analytical biochemistry 1986 May; 155(1):23-7/Name matches: sodium dodecyl sulfate coomassie brilliant blue r
Abstract:A simple method for the extraction of Coomassie brilliant blue R from stained protein bands excised from polyacrylamide gels is described. Spectrophotometric measurement of the eluted dye forms the basis of a sensitive assay to quantitate proteins in gels in the range 0.5-10 micrograms. The method requires no unusual equipment and is suitable for measurement of multiple samples. The polypeptide is not extracted and remains available for further analysis. The technique has been applied to three proteins and gels of various acrylamide percentages.

---

2.Preliminary Extraction and Identification of the 44.5 kDa Outer Membrane Proteins Isolated from Bovine Fusobacterium necrophorum (AB)/PMID 24426142; Indian journal of microbiology 2013 Dec; 53(4):395-9/Name matches: silver coomassie brilliant blue r-250
Abstract:Fusobacterium necrophorum (AB) in the pharynx, respiratory tract, female reproductive tract or urinary system is the causative agent of footrot and hepatic abscesses in animals and acute Lemierre's syndrome in humans. Current methods do not effectively protect animals and humans against F. necrophorum (AB). The outer membrane proteins (OMP) of F. necrophorum (AB) can be used as new material to protect against the diseases induced by F. necrophorum (AB). The aim of this study was to extract OMP and examine the immunogenic response of OMP. The preliminary extraction of OMP of F. necrophorum (AB) was identified by SDS-PAGE and stained by Coomassie Brilliant Blue R-250 (CB B R-250) and silver staining methods. The results showed that only a major band of 44.5 kDa was observed when staining the gel using CB B R-250. This band represented the target protein. In contrast, many small bands were observed by the silver staining method. The OMP also exhibited immune biological activities according to western blot analysis. The brightest band among the multi-banding observed was the OMP. Thus, the OMP was obtained and had immunogenic activity. The results provide a new direction to protect animals and humans against F. necrophorum (AB) in the clinical setting.

---

3.Stabilization of thin-layer agarose gels after isoelectric focusing with polyacrylamide enables reverse imidazole-zinc staining and facilitates two-dimensional gel electrophoresis/PMID 18607574; Analytical and bioanalytical chemistry 2008 Sep; 392(1-2):239-45/Name matches: silver coomassie brilliant blue r-250
Abstract:Large-pore-size agarose gels provide excellent resolving capacity for high molecular weight biomolecules. Thin-layer agarose isoelectric focusing (IEF) gels on polyester support films are especially useful for the separation of large proteins based on their pI in native conformation, but the method has suffered from the lack of detection methods compatible with agarose gels in hydrated form. Recently, an acrylamide copolymerization method was reported to enable mass-spectrometry-compatible silver staining and in-gel digestion of proteins. In this study, the method was further applied by demonstrating successful reverse imidazole-zinc staining of thin-layer agarose IEF gels copolymerized with acrylamide. The sensitivity of the reverse staining method on the composite gel at its best equaled the sensitivity of the traditional dried agarose silver staining method. Owing to the increased durability and reversible detection, the reverse-stained first-dimension gel could be conveniently prepared for the second-dimension sodium dodecyl sulfate polyacrylamide gel electrophoresis by reduction and alkylation. In addition, the micropreparative generation of tryptic peptides of Coomassie brilliant blue R-250 stained proteins in the composite gel is demonstrated.

---

4.Comparing standard and microwave assisted staining protocols for SDS-PAGE of glycoproteins followed by subsequent PMF with MALDI MS/PMID 19154801; Journal of proteomics 2009 May; 72(4):628-39/Name matches: silver cbb r250
Abstract:The detection of glycoproteins on SDS-PAGE gels is a very challenging task as glycan moieties can inhibit the protein-dye interaction or protein-silver reaction slowing down or even completely preventing the staining process. Additionally the applied staining procedure can influence the total number of detected peptides after in-gel digestion. Three in SDS-PAGE commonly used staining procedures (silver nitrate, CBB R250 and colloidal CBB G250) were evaluated in terms of duration, sensitivity and obtainable sequence coverage after PMF. The staining procedures were performed with and without the assistance of microwave irradiation. Microwave treatment resulted in comparable band intensities and sensitivities as obtained by the original staining protocols (limit of detection for microwave assisted silver staining: 50 ng), but staining duration was significantly reduced, to 30 min for silver nitrate and to 1.5 h for CBB and cCBB staining method. PMF analysis by MALDI mass spectrometry was not affected by the microwave treatment. It was found that the total number of detected tryptic peptides has increased when applying microwave irradiation during the staining process (average sequence coverage 31-56% and 76% for Avidin).

---

5.Quantitation of proteins bound to polyvinylidene difluoride membranes by elution of coomassie brilliant blue R-250/PMID 2281857; Analytical biochemistry 1990 Sep; 189(2):169-72/Name matches: sodium dodecyl sulfate coomassie brilliant blue r-250
Abstract:A rapid and simple method for the quantitation of stained proteins bound to polyvinylidene difluoride (PVDF) membranes via the elution of Coomassie brilliant blue R-250 is described. A mixture of standard proteins was resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroblotted onto PVDF membranes. Spectrophotometric analysis of dye eluted from protein bands in the range of 0.5-10 micrograms gave a linear change in the absorbance at 595 nm. Maximal absorbance readings were attained following 5 min of dye elution, and the readings remained unchanged for elution times up to 60 min. The method requires no unusual reagents or equipment, is suitable for the analysis of multiple samples, and does not consume the protein in the process of quantitation. This technique provides a useful means for the quantitation of proteins bound to PVDF membranes prior to amino acid sequence determination, immunological analysis, or other biochemical characterizations.