Far-western blotting

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Far-western blotting is a molecular biological method which is based on the technique of Western blotting. While usual western blotting uses an antibody to detect a protein of interest, far-western blotting uses a protein, which can bind the protein of interest. Thus, whereas western blotting is used for the detection of certain proteins, far-western blotting is rather employed to detect protein:protein interactions.

[edit] Method

In conventional Western blotting, gel electrophoresis is used to separate proteins from a sample; these proteins are then transferred to a membrane in a 'blotting' step. In a Western blot, specific proteins are then identified using an antibody probe.

Far-western blotting employs non-antibody-proteins – not antibodies – to probe the far-western blot. In this way, binding partners of the probe (or the blotted) protein may be identified.

The probe protein can then be identified through the usual methods—it may be radiolabelled; it may bear a specific affinity tag like His or FLAG; or it may be a protein for which a reliable antibody exists.