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Western Blot for weakly expressed protein - (Mar/25/2008 )

Hi, nice to meet you all here. Just a little question about Western Blot. In fact my protein is weakly expressed in the cell, is there any protocol or special kit for running a WB? Thanks. rolleyes.gif

-vincenttianfr-

Try immunoprecipitation and then western to detect. That'll enrich your protein.

-BioWizard v0.0.1-

you can amplify your detection by using a biotin-avidin (or streptavidin) system for your secondary antibody.

you have a biotinylated secondary and avidin (or streptavidin) and biotinylated detection enzyme (hrp or ap). this way you will have more than one enzyme molecule per antibody molecule.

-mdfenko-

QUOTE (vincenttianfr @ Mar 25 2008, 01:24 PM)
Hi, nice to meet you all here. Just a little question about Western Blot. In fact my protein is weakly expressed in the cell, is there any protocol or special kit for running a WB? Thanks. rolleyes.gif


in case your overexpressed protein carries a tag you may use it for specific enrichment before doing IB

-The Bearer-