Recombinant protein or a chemically synthesized bioactive fragment is immobilized on resin and used as a probe to capture interacting proteins directly from a cell extract. Affinity-purified proteins are fractionated by gel electrophoresis and visualized by Coomassie staining. Proteins that interact specifically are identified by comparing this gel profile to one obtained from cell lysates passed over a control resin lacking the immobilized probe protein. - [Read Affinity Purification of Interacting Proteins from Cell Lysates Protocol]
An excellent guide on the analysis of proteins on SDS-PAGE gels, through staining with coomassie blue dye and western blot analysis. Analysis of Protein Gels (SDS-PAGE). David R. Caprette, Rice University. - [Read Analysis of Protein Gels (SDS-PAGE)]
The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie Brilliant blue to protein. There is no interference from cations nor from carbohydrates such as sucrose.
Detergents such as sodium dodecyl sulfate and triton x-100 can interfere with the assay, as well as strongly alkaline solutions.
Includes a general overview of the procedure and preparation of the standards in the protocol. - [Read Bradford Assay Method]
Includes Abbreviations, Background, and Procedure steps using BSA. The Bradford protein assay (1) is one of several simple methods commonly used to determine the total protein concentration of a sample. The method is based on the proportional binding of the dye Coomassie to proteins. The assay is colorimetric; as the protein concentration increases, the color of the test sample becomes darker. Coomassie absorbs at 595 nm. - [Read Bradford Protein Concentration Assay]
Coomassie Blue Staining Protocol- http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/Proteomics/StainingProcedures.html#Coomassie
The cytotoxic effect of chemicals upon mammalian cells, such as BALB/c 3T3 and HepG2, in culture is measured by highest tolerated dose (HTD), cell viability (Neutral Red) and total cell protein (coomassie blue). - [Read Neutral Red Cytotoxicity Assay Protocol]
Protein Staining Protocols- http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/Proteomics/StainingProcedures.html
Protocols for protein staining. Includes: Coomassie Blue Staining; Silver Nitrate Staining; Zinc Imidazole Negative Staining. - [Read Protein Staining Protocols]
Colloidal coomassie blue staining, Destaining, Reduction/Alkylation, Peptide extraction For MALDI-MS analysis and For MALDI-MS analysis protocols and methods. Institut Curie Paris. - [Read Proteomic protocols and Peptide Extraction]