Staining and quantification of proteins separated by polyacrylamide gel electrophoresis

Syrový, I; Hodný, Zdeněk

doi:10.1016/0378-4347(91)80229-6

Cited by 75 publications

(46 citation statements)

References 96 publications

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“…It also belongs to the triphenylmethane dye family, but, unlike CBB R, it does not bind to ampholytes and can, therefore, be used as a stain for IEF gels also without separate protein fixation and ampholyte removal step. In quantitation, Fast Green proved slightly less sensitive than CBB, but showed a broader linear range (two orders of magnitude [32,33]). …”

Section: Fast Green Fcfmentioning

confidence: 99%

Protein stains for proteomic applications:Which, when, why?

Crawford²,

2006

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This review recollects literature data on sensitivity and dynamic range for the most commonly used colorimetric and fluorescent dyes for general protein staining, and summarizes procedures for the most common PTM-specific detection methods. It also compiles some important points to be considered in imaging and evaluation. In addition to theoretical considerations, examples are provided to illustrate differential staining of specific proteins with different detection methods. This includes a large body of original data on the comparative evaluation of several pre-and post-electrophoresis stains used in parallel on a single specimen, horse serum run in 2-DE (IPG-DALT). A number of proteins/protein spots are found to be over-or under-revealed with some of the staining procedures.

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Section: Fast Green Fcfmentioning

confidence: 99%

Protein stains for proteomic applications:Which, when, why?

Crawford²,

2006

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“…Although Coomassie brilliant blue stain is a reproducible, low-cost, and MS compatibility method, but it is relatively insensitive and time-consuming [2][3][4]. Silver staining is the most sensitive staining method for the visualization of protein in gels, up to 100 times more sensitive than Coomassie brilliant blue staining, but it lacks reproducibility and shows poor linearity with protein concentration [5][6][7][8][9].…”

Section: Introductionmentioning

confidence: 99%

High‐throughput negative detection of SDS‐PAGE separated proteins and its application for proteomics

et al. 2010

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A negative detection method for proteins on SDS-PAGE is described. In this method, Eosin Y (EY) was selectively precipitated in the gel background, which is absent from those zones where proteins are located through the formation of a stable water-soluble protein-dye complex. Negative staining of proteins using EY, allows high-sensitivity, lowcost, and simple protocol. The new described method takes less than an hour to complete all the protocol, with a detection limit of 0.5 ng of single protein band. Comparing with imidazole-zinc negative stain, EY dye provides broader linear dynamic range, higher sensitivity and reproducibility, and better obvious contrast between the protein bands or spots and background. Furthermore, the novel technique developed here presented a real practical method for simultaneous processing of multiple gels, which makes it possible to perform high-throughput staining for proteome research. Additionally, we have also compared the influence of staining method on the quality of mass spectra by PMF.

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“…We have compared Coomassie Brilliant Blue (CBB) G-250 and 35 S for protein detection. As it is commonly assumed that different proteins have different binding capacities for CBB G-250 [30][31][32][33][34], we studied the staining efficiency of CBB G-250 as a function of the proportion of the different amino acids in the protein. We also compared four software packages dedicated to image analysis: Melanie 3.0, Progenesis, ImageQuaNT and Kepler.…”

Section: Introductionmentioning

confidence: 99%

Assessing factors for reliable quantitative proteomics based on two‐dimensional gel electrophoresis

et al. 2004

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3IFR 87. La plante et son environnement. ISV, Gif-sur-Yvette, FranceWe statistically analysed various factors to get accurate estimates of protein quantities from two-dimensional gels. Yeast proteins were labelled with 35 S or stained with Coomassie Brilliant Blue G-250, and spots were automatically quantified with software packages Kepler, ImageQuaNT, Melanie 3.0 and Progenesis. The different software packages proved to have very similar performances. With 35 S-labelled actin spot as a reference, we studied the staining efficiency of colloidal Coomassie blue as a function of amino acid composition of the protein, and derived an equation to estimate the number of molecules per cell from blue-stained proteins. Absolute quantification of most glycolytic enzymes was carried out in two yeast strains.

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Staining and quantification of proteins separated by polyacrylamide gel electrophoresis

Cited by 75 publications

References 96 publications

Protein stains for proteomic applications:Which, when, why?

Protein stains for proteomic applications:Which, when, why?

High‐throughput negative detection of SDS‐PAGE separated proteins and its application for proteomics

Assessing factors for reliable quantitative proteomics based on two‐dimensional gel electrophoresis

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