Coomassie brilliant blue g 250 dye

Manufactured by Thermo Fisher Scientific

Sourced in United States

Coomassie Brilliant Blue G-250 Dye is a protein staining reagent used in various laboratory applications. It is a bright blue dye that binds to proteins, allowing for the visualization and quantification of proteins in samples.

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Lab products found in correlation

Tween 20, by Merck Group (2 mentions) Red blood cell lysis buffer, by Thermo Fisher Scientific (1 mentions) Ovalbumin (ova), by MP Biomedicals (1 mentions) Tris glycine sds electrophoresis buffer, by Bio-Rad (1 mentions) Carbamoylcholinechloride, by Bio-Techne (1 mentions) Phosphate citrate buffer, by Merck Group (1 mentions) Phosphate buffered saline (pbs), by Corning (1 mentions) Ms grade trypsin, by Promega (1 mentions) Silverquest silver staining kit, by Thermo Fisher Scientific (1 mentions) Captureselect fcxl affinity matrix, by Thermo Fisher Scientific (1 mentions) Laemmli sample buffer 4, by Bio-Rad (1 mentions) Pageruler plus prestained protein ladder, by Thermo Fisher Scientific (1 mentions) Polyvinylpyrrolidone, by Merck Group (1 mentions) Chemidoc mp, by Bio-Rad (1 mentions)

Close spelling variants of this product

Coomassie Brilliant Blue G-250 Coomassie blue G-250 dye Coomassie Brilliant Blue dye Coomassie Blue G-250 Brilliant Blue G-250 Coomassie Brilliant Blue Coomassie G-250 Coomassie Blue dye Coomassie blue

8 protocols using coomassie brilliant blue g 250 dye

Quantification of Intestinal Protein Content

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Protein content of the human intestinal extracts was estimated using Bradford protein assay according to the manufacturer’s instructions. This involved Bio-Rad based on the Coomassie Brilliant Blue G-250 dye (ThermoFisher Scientific, Hemel Hempstead, UK). The analysis was made in triplicate according to the manufacturer’s protocol using bovine serum albumin as a standard. To minimize the effect of RIPA buffer components on the determination of protein content in the total mucosal protein extracts, mucosal intestine samples were diluted 100 times in HPLC water before the assay was performed.

Couto N., Al-Majdoub Z.M., Gibson S., Davies P.J., Achour B., Harwood M.D., Carlson G., Barber J., Rostami-Hodjegan A, & Warhurst G. (2020). Quantitative Proteomics of Clinically Relevant Drug-Metabolizing Enzymes and Drug Transporters and Their Intercorrelations in the Human Small Intestine. Drug Metabolism and Disposition, 48(4), 245-254.

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Native Polyacrylamide Gel Electrophoresis

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Native gel electrophoresis was set up in a two‐phase polyacrylamide gel. Acrylamide concentration was 4% in the stacking gel (pH = 6.8) and 10% in the resolving gel (pH = 8.8). After 30‐min pre‐electrophoresis without sample addition at constant 100 V on ice, the electrophoresis was performed for another 2.25 h at 200 V in native ‘ELFO buffer’ (30.3 g·L⁻¹ TRIS base, 144 g·L⁻¹ glycine, pH = 8.7). During electrophoresis, the whole apparatus was placed on ice. The gel was stained with Coomassie Brilliant Blue G250 dye (Thermo Fisher Scientific, Waltham, MA, USA).

Benedek A., Pölöskei I., Ozohanics O., Vékey K, & Vértessy B.G. (2017). The Stl repressor from Staphylococcus aureus is an efficient inhibitor of the eukaryotic fruitfly dUTPase. FEBS Open Bio, 8(2), 158-167.

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Quantifying p53 Levels in HeLa Cells

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The P53 content of HeLa cells was measured, using Human P53 ELISA Kit (Diaclone, France. Cat#: 850.630.048). Seventy-two hours' post-transfection, the transfected cells were lysed and their protein concentration was quantified using Coomassie Brilliant Blue G-250 Dye (Thermo Scientific, USA). One hundred µl of each sample was used for P53 ELISA in a duplicate array. ELISA was performed according to the manufacturer's instructions. P53 concentrations were normalized and reported as percentage of untreated control cells for each sample.

Pirouzfar M., Amiri F., Dianatpour M, & Takhshid M.A. (2020). CRISPR/Cas9-mediated knockout of MLL5 enhances apoptotic effect of cisplatin in HeLa cells in vitro. EXCLI Journal, 19, 170-182.

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Palmitoylation Assay for YKT6

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The palmitoylation of YKT6 was determined using the palmitoylation assay as previously described [35 (link)]. Samples were tested by SDS/PAGE, and the gels were stained with Coomassie Brilliant Blue G-250 Dye (Thermo Fisher Scientific, Waltham, MA, USA).

Sun C., Wang P., Dong W., Liu H., Sun J, & Zhao L. (2020). LncRNA PVT1 promotes exosome secretion through YKT6, RAB7, and VAMP3 in pancreatic cancer. Aging (Albany NY), 12(11), 10427-10440.

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Pollen Extraction and Immunoassay Protocol

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Common RW (Ambrosia elatior) pollen was obtained from Pharmallerga (Lisov, Czech Republic). All chemicals and materials were purchased from Fisher Scientific (PA, USA) unless otherwise stated. Tween 20 and phosphate-citrate buffer tablets were purchased from Sigma-Aldrich. Phosphate-buffered saline (PBS) was bought from Mediatech, Inc. (Manassas, VA, USA). All cell culture reagents, lane marker sample reducing buffer, Coomassie brilliant blue G-250 dye, fluorescein isothiocyanate (FITC)-conjugated OVA, and O-phenylenediamine (OPD) tablets were purchased from Thermo Fisher Scientific (Waltham, MA, USA). OVA was purchased from MP Biomedicals (Solon, OH, USA). Non-fat dry milk, 4–20% mini-PROTEAN TGX precast gels for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Tris/Glycine/SDS electrophoresis buffer, and protein standards were obtained from Bio-Rad Laboratories, Inc. (Hercules, CA, USA). Carbamoylcholine chloride was obtained from Tocris Bioscience (Bio-Techne, Minneapolis, MN, USA). Horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG, IgG1, IgG2a, IgA, and IgE antibodies were bought from Southern Biotech (Birmingham, AL, USA). Common RW extract was obtained from Greer® (Lenoir, NC, USA). Red blood cell lysis buffer was purchased from eBioscience, Inc. (San Diego, CA, USA).

Uddin M.J, & Gill H.S. (2018). From allergen to oral vaccine carrier: a new face of ragweed pollen. International journal of pharmaceutics, 545(1-2), 286-294.

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Coomassie Gel-Based Protein Digestion Protocol

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After electrophoretic protein separation, gels were stained with Coomassie Brilliant Blue G-250 dye (ThermoFisher) for 2 h and left to destain overnight in the destain solution (10% acetic acid/20% methanol). The destained gel pieces were incised, washed with HPLC water and dehydrated with neat acetonitrile (ACN). Proteins were reduced with 2 mM Tris(2-carboxyethyl)phosphine (TCEP) in 50 mM ammonium bicarbonate (NH₄HCO₃, AmBic) for 1 h at 37°C and then dehydrated with ACN. The reduced proteins were alkylated with 50 mM iodoacetamide (IAA)/50 mM AmBic, for 20 min in dark with rotation. The gel pieces were dehydrated again with ACN to remove all the reagents. MS-grade trypsin (10 ng/μL) (Promega) was added to the samples and incubated for 30 min on ice. After the excess of trypsin was removed from tubes, 25 mM AmBic was added to cover the gel pieces and incubate overnight at 37°C. Digested peptides were then extracted from the gel with 50% ACN/0.1% trifluoroacetic acid solution. Samples were dried in a SpeedVac, re-dissolved in 15 μL 0.1% formic acid and submitted for LC-MS/MS analysis.

Lagundžin D., Hu W.F., Law H.C., Krieger K.L., Qiao F., Clement E.J., Drincic A.T., Nedić O., Naldrett M.J., Alvarez S, & Woods N.T. (2019). Delineating the role of FANCA in glucose-stimulated insulin secretion in β cells through its protein interactome. PLoS ONE, 14(8), e0220568.

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Capture and Analyze IgG-BCRs from Ramos B Cells

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To capture IgG-BCRs, 20 million Ramos B cells were washed with PBS, to remove FCS from the cell culture medium, and lysed in 8 ml PBS + 1% Triton-X100 for 60 min at 37 °C. IgG-BCRs were captured from cell lysis supernatants using 20 µl CaptureSelect^TM FcXL Affinity Matrix (Thermo Fisher Scientific) and an overnight incubation at 4 °C. B cell secreted IgG were captured from 12 ml Ramos B cell supernatant (cultured at a density of two million cells/ml) by a 4 °C overnight incubation with 20 µl CaptureSelect^TM FcXL Affinity Matrix (Thermo Fisher Scientific). Laemmli sample buffer (4×) (Bio Rad) was added to the IgG/FcXL bead slurry, boiled for 5 min at 95 °C, and loaded on a 4 to 15% SDS gel (Bio Rad). Proteins were detected with Coomassie Brilliant Blue G-250 Dye (Thermo Fisher Scientific) or the SilverQuest^TM Silver Staining Kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. The size was determined using the PageRuler^TM Plus Prestained Protein Ladder (Thermo Fisher Scientific).

Kissel T., Derksen V.F., Bentlage A.E., Koeleman C., Hafkenscheid L., van der Woude D., Wuhrer M., Vidarsson G, & Toes R.E. (2024). N-linked Fc glycosylation is not required for IgG-B-cell receptor function in a GC-derived B-cell line. Nature Communications, 15, 393.

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SDS-PAGE and Lectin Blot Analysis

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Aliquots of 75 µg PRM were subjected to denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using 12% (w/v) polyacrylamide gels and separated at 100 V for 90 min. The separated material was either used for conventional staining with Coomassie Brilliant Blue G-250 Dye (Thermo Fisher Scientific) or transferred to a nitrocellulose membrane for blotting. The membrane was blocked with 5% (w/v) polyvinylpyrrolidone (Sigma-Aldrich) in TBS buffer (50 mM Tris-HCl, pH 7.6, 150 mM NaCl) for 60 min, then was incubated for 2 h at room temperature with 1 mg mL⁻¹ ConA-fluorescein (FITC; Sigma-Aldrich) in TBS added with 0.1% (v/v) Tween 20 (Sigma-Aldrich). Then, the membrane was washed six times with TBS-Tween and inspected in a ChemiDoc MP (Bio-Rad), using a wavelength of 520 nm.

García-Carnero L.C., Salinas-Marín R., Lozoya-Pérez N.E., Wrobel K., Wrobel K., Martínez-Duncker I., Niño-Vega G.A, & Mora-Montes H.M. (2021). The Heat Shock Protein 60 and Pap1 Participate in the Sporothrix schenckii-Host Interaction. Journal of Fungi, 7(11), 960.

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