Coomassie stain solution

Manufactured by Bio-Rad

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Coomassie stain solution is a laboratory reagent used for the detection and quantification of proteins in various applications, such as gel electrophoresis and western blotting. The solution contains Coomassie Brilliant Blue dye, which binds to proteins and produces a colored complex, allowing for the visualization and analysis of protein samples.

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Coomassie stain (22 citations) Coomassie blue stain solution (2 citations) Bio-Safe Coomassie Stain solution (2 citations)

4 protocols using coomassie stain solution

Exosome Proteome Profiling by MMP14 Cleavage

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One hundred micrograms of exosomes of each type were incubated with or without 0.2 μg of the catalytic domain of MMP14 enzyme (Calbiochem) in 40 μL of MMP reaction buffer for 1 hour at 37°C. The MMP14 enzyme reaction was terminated by adding 4X LDS sample buffer, including β-mercaptoethanol. Exosomal proteins were separated by electrophoresis using 4% to 20% polyacrylamide gel (Invitrogen) and stained by Coomassie stain solution (Bio-Rad). For mass spectrometry, proteins bands were cut off from stained SDS-PAGE gel with Coomassie blue (Bio-Rad). Samples were analyzed by the UCSD Proteomics Facility (http://bpmsf.ucsd.edu). Counts for the total spectrum from Scaffold 4 viewer software, and molecular weight and accession numbers were compared.

Han K.Y., Chang J.H, & Azar D.T. (2019). MMP14-Containing Exosomes Cleave VEGFR1 and Promote VEGFA-Induced Migration and Proliferation of Vascular Endothelial Cells. Investigative Ophthalmology & Visual Science, 60(6), 2321-2329.

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Gelatin Zymography Assay for MMPs

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Conditioned media were obtained after a 30-h incubation of the different BGC823 and MGC803 cells in serum-free medium and were then concentrated 80-fold using Amicon Ultra Centrifugal Filter Units (Millipore) and normalized by protein concentration. Samples were loaded on 10% SDS-PAGE gels containing 0.1% gelatin. Electrophoresis was carried out under nonreducing conditions at 100 V and 4°C. The gels were then washed in 2.5% Triton X-100, incubated in substrate buffer (50 mmol/L Tris–HCl, pH 8.0; 50 mmol/L NaCl; 10 mmol/L CaCl₂; and 0.05% Brij 35) for 40 h at 37°C, stained with Coomassie stain solution (Bio-Rad), and destained in 20% methanol and 10% acetic acid. Gelatinolytic activity was identified as a clear band on a blue background. The activities of secreted MMPs were detected using gelatin zymography as previously described [36 (link)], with several modifications.

Xu T.P., Huang M.D., Xia R., Liu X.X., Sun M., Yin L., Chen W.M., Han L., Zhang E.B., Kong R., De W, & Shu Y.Q. (2014). Decreased expression of the long non-coding RNA FENDRR is associated with poor prognosis in gastric cancer and FENDRR regulates gastric cancer cell metastasis by affecting fibronectin1 expression. Journal of Hematology & Oncology, 7, 63.

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Estradiol-Mediated Ubiquitination Assay

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17β-estradiol was purchased from Sigma-Aldrich (St. Louis, MO, USA), MG-132 from Enzo Life Sciences (Farmingdale, NY, USA), Cy5-conjugated Streptavidin from Jackson ImmunoResearch Laboratories (West Grove, PA, USA), and Coomassie Stain Solution from Bio-Rad (Hercules, CA, USA). The recombinant KAT7 (Hbo1 purified from Sf9 cells), MurF1, GST-tagged p53, and ubiquitin proteins were purchased from Active Motif (Carlsbad, CA, USA), R&D systems (Minneapolis, MN, USA), EMD Millipore (Temecula, CA, USA) and UBPBio (Aurora, CO, USA), respectively. Anti-Hbo1 antibody was described.^{17 (link))} The following commercial antibodies were purchased: anti-FLAG monoclonal and anti-α-tubulin monoclonal from Sigma-Aldrich (St. Louis, MO, USA), anti-HA monoclonal from Roche (Basel, Switzerland), anti-ERα polyclonal from Santa Cruz (Dallas, TX, USA), anti-ubiquitin polyclonal from Cell Signaling Technology (Danvers, MA, USA), and anti-GST polyclonal and anti-His-tag monoclonal from Medical and Biological Laboratories (Nagoya, Japan).

IIZUKA M., SUSA T., TAMAMORI-ADACHI M., OKINAGA H, & OKAZAKI T. (2017). Intrinsic ubiquitin E3 ligase activity of histone acetyltransferase Hbo1 for estrogen receptor α. Proceedings of the Japan Academy. Series B, Physical and Biological Sciences, 93(7), 498-510.

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Artesunate Derivatization and Characterization

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Artesunate, dihydroartemisinin (DHA), bromotrimethylsilane (TMBS), sodium azide (NaN₃), tetrahydrofuran (THF), triphenyl phosphine (PPh₃), 1-hydroxybenzotriazole hydrate, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, acetyl chloride, N,N-dimethylformamide (DMF), dimethyl-sulfoxide (DMSO), dichloromethane (CH₂Cl₂), hydrogen peroxide (H₂O₂), and Protease cocktail were purchased from Sigma-Aldrich (St. Louis, MO, USA). Imm. Drystrip pH 3–11 (Cat No. 17600377) and buffer (Cat No. 17600440) purchased from GE. RIPA lysis and extraction buffer was purchased from Pierce Biotechnology (Cat. No. 89900). Coomassie stain solution is from Bio-Rad (Cat. No. 161-0436). Nrf2 and corresponding secondary antibodies are from Santa Cruz Biotechnology (Cat. No. sc-722, sc-2030), Keap1 antibody is from Cell Signalling Technology (Product No. 4617S, 8047S), ECL reagent is from GE Healthcare (Cat. No. RPN2232), UltraLink Immobilized Streptavidin beads are from Thermo Scientific (Cat. No. 20349), Centrifuge Columns are from Pierce (Cat. No. 89896), OMIX tips are from Agilent technologies (Part No. A57003100). Unless otherwise noted, all the materials were obtained from commercially available sources and were used without further purification.

Ravindra K.C., Ho W.E., Cheng C., Godoy L.C., Wishnok J.S., Ong C.N., Wong W.S., Wogan G.N, & Tannenbaum S.R. (2015). An Untargeted Proteomics and Systems-based Mechanistic Investigation of Artesunate in Human Bronchial Epithelial Cells. Chemical research in toxicology, 28(10), 1903-1913.

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