Quantification of attached cells in microtiter plates based on Coomassie brilliant blue G-250 staining of total cellular protein

Laughton, Craig W.

doi:10.1016/0003-2697(84)90187-8

Cited by 55 publications

(18 citation statements)

References 7 publications

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“…This parallels the findings of Barer et al (1986) and Laughton (1984) who pointed out that in each cell system, staining kinetics should be estimated. In the system employed in this study with BHK cells, exposure to Coomassie blue for 30 rnin gave a suitable range of absorbance values.…”

Section: Discussionsupporting

confidence: 88%

“…Adherent cells have been quantitated by several methods including direct microscopy (Patterson, 1979), assay of cell enzymes (Connolly et al, 1986) or estimation of cell protein (Laughton, 1984;Shopsis and Mackay, 1984;Baumgarten, 1985). One method (Shopsis and Mackay, 1984) required initial lysis of cells before Lowry micro-assay of cell protein.…”

Section: Discussionmentioning

confidence: 99%

“…One method (Shopsis and Mackay, 1984) required initial lysis of cells before Lowry micro-assay of cell protein. Other Coomassie blue protein dye-binding assays have been described (Laughton, 1984;Baumgarten, 1985). These used a modification of the Bradford protein assay (Bradford, 1976), which depends on the colour change of Coomassie blue G-250 at low pH from brown-red to blue when bound to protein.…”

Section: Discussionmentioning

confidence: 99%

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Quantitative cell-adhesion assay for Clostridium difficile cytotoxin

Mayall

Edmonds²,

Griffin³

1987

Journal of Medical Microbiology

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Summary.A quantitative assay for Clostridium dzficile cytotoxin has been developed, based on the observation that suspended fibroblasts exposed to cytotoxin fail to adhere to plastic. A dye-binding technique was used to quantitate adherent cells, in order to obviate microscopy. Adherent BHK cells were fixed with glutaraldehyde and cell protein was stained with Coomassie blue R-250. Cell-bound dye was eluted and estimated spectrophotometrically. The amount of eluted dye was proportional to the number of adherent cells and cell staining was time dependent. Cytotoxin was purified by gel-permeation and ion-exchange chromatography and migrated as a single band on SDS-PAGE. After exposure of suspended BHK cells to purified cytotoxin, their adhesion to plastic was inhibited in a manner which depended on concentration of cytotoxin and on time and temperature of exposure. This study provides the basis for a C . dzficile cytotoxin assay that is quantitative, rapid and reproducible and may have wider applicability in the study of other toxins or agents that inhibit cell adhesion.

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Section: Discussionsupporting

confidence: 88%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Quantitative cell-adhesion assay for Clostridium difficile cytotoxin

Mayall

Edmonds²,

Griffin³

1987

Journal of Medical Microbiology

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“…Staining methods include silver staining (7), Coomassie Brilliant Blue staining (10), fluorescent staining (3) etc. The color indicates the percentages of proteins, while the strength provides the information of amount.…”

Section: Introductionmentioning

confidence: 99%

A Label-Free Strategy for both Qualification and Quantitation of Protein Based on Tandem Mass Spectrometry

Wang

Zhao

Liu

et al. 2012

Biotechnology & Biotechnological Equipment

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“…To overcome these problems, a great variety of workable colorimetric assays for cell quantitation has been developed in the last years. Widely used techniques include staining with neutral red (12,13), crystal violet (14 -16), and Coomassie blue (17,18) as well as procedures based on methylene blue (19,20), naphthol blue-black (21), naphthol yellow S (22), MayGrü nwald-Giemsa (23), and sulforhodamine B (6). These assays mainly focus on the measurement of cellular parameters such as protein (5,6), DNA (7), or whole macromolecular contents (8), take advantage of accumulation of dyes in cell compartments (9 -11), or represent cell nuclei or simple biomass stains (e.g., 6).…”

mentioning

confidence: 99%