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