Abstract
Most traditional post-electrophoretic processes need several hours to several days to finish the whole staining process and traditional staining solutions all contain methanol, acetic acid, or phosphoric acid, which not only produce the unpleasant smell but also cause environmental pollution. Here a fixation-free, fast protein staining method in sodium dodecyl sulfate–polyacrylamide gel electrophoresis using Coomassie blue is described. The protocol includes only staining and quick washing steps, can be completed in 0.5 h. It has a sensitivity of 10 ng. In addition, the dye stain does not contain any acid or methanol.
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References
Walker J (ed) (2002) The protein protocols handbook, 2nd edn. Humana Press, Totowa, pp 61–67
Shofuda K, Nagashima Y, Kawahara K, Yasumitsu H, Miki K et al (1998) Elevated expression of membrane-type 1 and 3 matrix metalloproteinases in rat vascular smooth muscle cells activated by arterial injury. Lab Investig 78:915–923
Kawsar S, Fujii Y, Matsumoto R, Ichikawa T, Tateno H et al (2008) Isolation, purification, characterization and glycan-binding profile of a d-galactoside specific lectin from the marine sponge, Halichondria okadai. Comp Biochem Physiol B Biochem Mol Biol 150:349–357
Kawsar S, Takeuchi T, Kasai K, Fujii Y, Matsumoto R et al (2009) Glycan-binding profile of a D-galactose binding lectin purified from the annelid, Perinereis nuntia ver. vallata. Comp Biochem Physiol B Biochem Mol Biol 152:382–389
Yasumitsu H, Ozeki Y, Kawasar SM, Fujii Y, Sakagami M et al (2010) RAMA stain: a fast, sensitive and less protein-modifying CBB R250 stain. Electrophoresis 31:1913–1917
Berggren KN, Schulenberg B, Lopez MF, Steinberg TH, Bogdanova A et al (2002) An improved formulation of SYPRO Ruby protein gel stain: comparison with the original formulation and with a ruthenium II tris (bathophenanthroline disulfonate) formulation. Proteomics 2:486–498
Candiano G, Bruschi M, Musante L, Santucci L, Ghiggeri M et al (2004) Bluesilver: a very sensitive colloidal Coomassie G-250 staining for proteome analysis. Electrophoresis 25:1327–1333
Fazekas D, Groth S, Webster G, Datyner A (1963) Two new staining procedures for quantitative estimation of proteins on electrophoretic strips. Biochim Biophys Acta 71:377–391
Merril C, Goldman D, Sedman S, Ebert M (1980) Ultrasensitive stain for proteins in polyacrylamide gels shows reqional variation in cerebrospinal fluid proteins. Science 211:1437–1438
Weber K, Osborn M (1969) The reliability of molecular weight determinations by dodecyl sulfatepolyacrylamide gel electrophoresis. Bid Chem 244:4406–4412
Bennet J, Scott J (1971) Quantitative staining of fraction 1 protein in polyacrylamide gels using Coomassie brilliant blue. Anal Biochem 43:173–182
Bertolini M, Tankersley L, Schroeder D (1976) Staining and destaining polyacrylamide gels: a comparison of coomassie blue and fast green protein dyes. Anal Biochem 71:6–13
Kalivarathan D, Vijayan S, Sridhar B, Krishnamurthy S, Pennathur G (2011) In-gel staining of proteins in native poly acryl amide gel electrophoresis using tetrakis(4-sulfonato phenyl)porphyrin. Anal Sci 27:101–103
Pink M, Verma N, Rettenmeier AW, Schmitz-Spanke S (2010) CBB staining protocol with higher sensitivity and mass spectrometric compatibility. Electrophoresis 31:593–598
Jung C, Sang Y, Hee H, Dong C, Gyurng Y (1996) A modified Coomassie blue staining of proteins in polyacrylamide gels with Bismark brown R. Anal Biochem 236:82–84
Acknowledgments
This work was supported by the grants from National Natural Science Foundation of China (No. 81673337).
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Dong, Wh., Wang, F., Zhang, Jh., Zhou, Ys., Zhang, Ly., Wang, Ty. (2018). A Simple, Time-Saving Dye Staining of Proteins in Sodium Dodecyl Sulfate–Polyacrylamide Gel Using Coomassie Blue. In: Kurien, B., Scofield, R. (eds) Protein Gel Detection and Imaging. Methods in Molecular Biology, vol 1853. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8745-0_5
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DOI: https://doi.org/10.1007/978-1-4939-8745-0_5
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