Far-Western Blot Analysis

Far-WesternBlot is a molecular biological method, which is based on the technique of Westernblot, to detect in vitro protein-protein interactions, especially for detecting interactions that do not require the native structure of the protein of interest. While conventional Westernblot utilizes an antibody to detect a protein of interest, a non-antibody protein, which can bind the protein of interest, takes the place of antibody in far-WesternBlot.

Far-WesternBlot employs non-antibody proteins to probe the proteins of interest and the bound partners may be identified with mass spec analysis. Prey proteins in a cell lysate are firstly separated by SDS/Native PAGE, and transferred to a membrane; the proteins after denatured and renatured are then blocked and probed, usually with purified bait proteins. The bait proteins are detected when incubating with bait proteins, if the bait proteins and the prey protein can form a complex.

In far-WesternBlot, multiple approaches can be used to detect protein-protein interactions, including:

Direct detection of prey protein with a radioactive bait protein;

Indirect detection with an antibody specific to the bait protein;

Indirect detection with an antibody specific to the tag of a fusion-tagged bait protein;

Indirect detection with a biotinylated bait protein and enzyme-conjugated avidin or streptavidin.

Creative Proteomics can provide far-WesternBlot service as you require. After a tech conversation with you about the experimental details, our tech team can provide customer-tailored services.

http://www.creative-proteomics.com/services/far-westernblot-analysis.htm