Coomassie brilliant blue staining

Manufactured by Thermo Fisher Scientific

Sourced in United States

Coomassie Brilliant Blue is a dye commonly used in protein staining for laboratory applications. It binds to proteins, allowing for their visualization and quantification. The stain is a simple, effective tool for analyzing protein samples in various biochemical and analytical procedures.

Automatically generated - may contain errors

Lab products found in correlation

Protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions) Pierce protein g agarose resin, by Thermo Fisher Scientific (1 mentions) Bolt bis tris plus gel, by Thermo Fisher Scientific (1 mentions) Freestyle 293 expression medium, by Thermo Fisher Scientific (1 mentions) Nitrocellulose membrane, by Bio-Rad (1 mentions) Goat anti rabbit igg peroxidase secondary antibody, by Merck Group (1 mentions) Pbs t, by Bio-Rad (1 mentions) Nupage 4 12 bis tris protein gel, by Thermo Fisher Scientific (1 mentions) Chemidoc mp imager, by Bio-Rad (1 mentions) Pbs t, by Merck Group (1 mentions) Image lab software, by Bio-Rad (1 mentions) Trans blot turbo system, by Bio-Rad (1 mentions) Clarity western ecl substrate, by Bio-Rad (1 mentions) 3xflag peptide, by Merck Group (1 mentions) Ez view red protein g beads, by Merck Group (1 mentions) Ezview red anti flag m2 agarose, by Merck Group (1 mentions) Halt phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Nupage 4 12 bis tris, by Thermo Fisher Scientific (1 mentions) Endo h, by New England Biolabs (1 mentions)

Close spelling variants of this product

Coomassie Brilliant Blue Coomassie blue staining Coomassie staining Coomassie blue

7 protocols using coomassie brilliant blue staining

Recombinant Antibody Production and Purification

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 3

HEK 293 cells and FreeStyle™ 293 Expression Medium (Invitrogen, USA) were used for recombinant antibody production. Transient transfection was done according to the manufacturer's instruction (Invitrogen, USA). To purify antibodies, the culture supernatant was applied to Pierce protein G agarose resin (ThermoFisher, USA) and were dialyzed against PBS. Purified antibody was analyzed by SDS-PAGE on 4-12% Bolt Bis-Tris plus gel (Invitrogen, USA) under reducing or non-reducing condition, and visualized by Coomassie brilliant blue staining (Invitrogen, USA).

Results from FIG. 1 showed all IgG4-reformatted anti-CD47 antibodies were properly expressed and formed. And also, greater than 90% purity of each individual antibody was obtained using Protein-G chromatography.

US11390677B2. Human anti-CD47 antibodies and uses thereof (2022-07-19). Trican Biotechnology Co., Ltd [TW]. Inventors: Huang-Tsu Chen [US], Jiun-Shyang Leou [TW], Chung-Yuan Hsu [TW], Cheng-Ke Li [TW], Yun-Ting Wang [TW], Li-Tsen Lin [TW], Shiou-Ting Chen [TW], Chen-Wei Chiang [TW].

+ Open protocol

+ Expand

Labeling and Characterizing Monoclonal Antibodies

Check if the same lab product or an alternative is used in the 5 most similar protocols

All human monoclonal antibodies were labeled with the IRDye 800 CW protein labeling kit following the manufacturer’s instructions (Microscale, Invitrogen). Briefly, the pH level was adjusted to 8.5 by adding 1 M potassium phosphate buffer to 200 µl of antibody solution (1 mg/ml of IgG in PBS). 1.4 µl of dissolved dye was added to the antibody solution and the mixture was incubated for 2 h at room temperature. Subsequently, free dye was removed using Zeba Spin Desalting columns according to the manufacturer’s instructions. Centrifuge columns were washed in PBS three times, and excess wash solution was removed by dry spin at 1,500 g. The columns were placed in a new collection tube and 100 µl of the sample was applied to each column. The tube was centrifuged at 1,500 g for 2 min to collect the sample. Integrity of labeled antibodies was confirmed using a non-reducing 12% Bis-Tris Protein Gel (NuPage, Invitrogen) according to manufacturer’s instruction. Gel was stained with Coomassie Brilliant Blue Staining (Invitrogen).
To confirm intensity of labeling, dot blots were performed with serial dilutions of infrared labeled antibodies, on a nitrocellulose membrane (Biorad, 0.45 µm), drying the membrane and scanning for signal intensity. Starting dilution was 20 µg/ml of infrared labeled antibody. Membrane analysis was performed using the Odysey infrared imaging system (LICOR).

Mader S., Brimberg L., Soltys J.N., Bennett J.L, & Diamond B. (2018). Mutations of Recombinant Aquaporin-4 Antibody in the Fc Domain Can Impair Complement-Dependent Cellular Cytotoxicity and Transplacental Transport. Frontiers in Immunology, 9, 1599.

+ Open protocol

+ Expand

Rab8a Protein Identification by SDS-PAGE

Check if the same lab product or an alternative is used in the 5 most similar protocols

Purified wildtype and mutant Rab8a proteins were separated on 4–20% SDS-PAGE, stained with Coomassie brilliant blue staining (Thermo Scientific) and a band corresponding to a ~ 50 kDa protein was excised. Protein identification was performed using MASCOT.

Madero-Pérez J., Fdez E., Fernández B., Lara Ordóñez A.J., Blanca Ramírez M., Gómez-Suaga P., Waschbüsch D., Lobbestael E., Baekelandt V., Nairn A.C., Ruiz-Martínez J., Aiastui A., López de Munain A., Lis P., Comptdaer T., Taymans J.M., Chartier-Harlin M.C., Beilina A., Gonnelli A., Cookson M.R., Greggio E, & Hilfiker S. (2018). Parkinson disease-associated mutations in LRRK2 cause centrosomal defects via Rab8a phosphorylation. Molecular Neurodegeneration, 13, 3.

+ Open protocol

+ Expand

Analyzing Ubiquitinated Proteins in B. divergens

Check if the same lab product or an alternative is used in the 5 most similar protocols

Reducing SDS PAGE and Western blot analyses were performed as described previously (Perner et al., 2016 ). Briefly, B. divergens lysates were analysed by SDS-PAGE using the NuPAGE 4–12% Bis-Tris Protein Gels and visualization by Coomassie Brilliant Blue staining (Thermofisher). For Western blot the unstained SDS PAGE gel loads were electro blotted to a PVDF (polyvinylidene difluoride) membrane using the Trans-Blot Turbo system (BioRad, USA). The membrane was blocked in 3% (w/v) non-fat skimmed milk in 1 × PBS with 0.05% Tween 20 (PBS-T; Sigma-Aldrich, USA). For immunostaining the membrane was first exposed to Anti-Ubiquitin K48 Linkage antibodies (Boston Biochem, USA) diluted by 1:2000 in PBS-T, washed (3 × 5 min) in PBS-T, and subsequently exposed to the goat anti-rabbit IgG-peroxidase secondary antibody (Sigma) diluted by 1: 2000 in PBS-T. After the final wash (4 × 5 min in PBS-T) the membrane was developed using the ClarityWestern ECL substrate (BioRad, USA), visualized in the ChemiDoc MP imager and analysed using Image Lab Software (BioRad, USA).

Jalovecka M., Hartmann D., Miyamoto Y., Eckmann L., Hajdusek O., O'Donoghue A.J, & Sojka D. (2018). Validation of Babesia proteasome as a drug target. International Journal for Parasitology: Drugs and Drug Resistance, 8(3), 394-402.

+ Open protocol

+ Expand

Isolation of FLAG-tagged Proteins for Proteomic Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

HEK293T cells were transfected with 3xFLAG tagged DAPK1, LRRK1, LRRK2, MASL1, or GFP plasmids using PEI reagent, collected 24 h after transfection and cells were lysed in the buffer: 20 mM Tris (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1% Triton, 10% Glycerol, protease inhibitor cocktail (Roche), and 1x Halt phosphatase inhibitor cocktail (Thermo Scientific). Lysates were precleared by centrifugation at 20 000 × g for 10 min and incubated for 1 h at 4 °C with EZview Red Protein G beads (Sigma) to remove proteins non‐specifically binding to agarose. After preclear with protein G beads, lysates were incubated for 1 h at 4 °C with EZview Red Anti‐FLAG M2 Agarose (Sigma) that is suitable for the immunoprecipitation of FLAG fusion proteins. Beads were washed six times with the wash buffer: 20 mM Tris (pH 7.5), 400 mM NaCl, 1% Triton and proteins were eluted in 25 mM Tris (pH 7.5), 150 mM NaCl, and 100 μg mL⁻¹ 3xFLAG peptide (Sigma). Protein yields and purity were estimated by staining gels with Coomassie brilliant blue staining (Thermo Scientific, Figure 1, Supporting Information).

Tomkins J.E., Dihanich S., Beilina A., Ferrari R., Ilacqua N., Cookson M.R., Lewis P.A, & Manzoni C. (2018). Comparative Protein Interaction Network Analysis Identifies Shared and Distinct Functions for the Human ROCO Proteins. Proteomics, 18(10), 1700444.

+ Open protocol

+ Expand

Deglycosylation of SWO1 Protein Using Endo H

Check if the same lab product or an alternative is used in the 5 most similar protocols

Deglycosylation of SWO1 was performed according to a standard Endo H protocol (New England Biolabs, Frankfurt, Germany). The purified protein (10 µg) was mixed with 10× denaturation buffer (5 % SDS, 400 mM DTT) up to 40 µL of total volume and incubated at 95 °C for 10 min. Subsequently, one-tenth volume of 10× G7 reaction buffer (500 mM sodium phosphate buffer, pH 7.5) was added. The mixture was incubated with 300 U Endo H at 37 °C for 90 min. Reaction was stopped by heating for 10 min to 95 °C. Deglycosylated samples and untreated controls were analyzed by SDS-PAGE (NuPAGE^® Bis–Tris 4–12 %) with glycostaining and subsequent Coomassie Brilliant Blue staining (Thermo Fisher Scientific, Waltham, MA, USA). Glycostaining was done with the Pro-Q Emerald 300 glycostain kit (Invitrogen, Carlsbad, CA) according the manufacturer’s protocol.

Eibinger M., Sigl K., Sattelkow J., Ganner T., Ramoni J., Seiboth B., Plank H, & Nidetzky B. (2016). Functional characterization of the native swollenin from Trichoderma reesei: study of its possible role as C1 factor of enzymatic lignocellulose conversion. Biotechnology for Biofuels, 9(1), 178.

+ Open protocol

+ Expand

Caspase-Mediated Cleavage of MVP

Check if the same lab product or an alternative is used in the 5 most similar protocols

For cleavage by the p20/p10 tetramers of active caspase, 3.5 mg of recombinant amino-terminal GST-tagged MVP (Abnova H00009961-P01, Taiwan) were incubated with 1 unit of caspase-1, -3, -7, -9, -8, -10, -2, and -6 in a 35 ml reaction containing 50 mM HEPES (pH 7.5), 3 mM EDTA, 150 mM NaCl, 0.005% (v/v) Tween-20, and 10 mM DTT. The reaction was incubated for 5 hours at 37 C. Reaction samples were separated on SDS-PAGE gels and cleavage of MVP was examined by Coomassie Brilliant Blue staining (Thermo Fisher Scientific) and immunoblotting.

Grossi S., Fenini G., Kockmann T., Hennig P., Di Filippo M, & Beer H.D. (2020). Inactivation of the Cytoprotective Major Vault Protein by Caspase-1 and -9 in Epithelial Cells during Apoptosis. The Journal of investigative dermatology, 140(7).

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!