RNAi-based validation of antibodies for reverse phase protein arrays.

ABSTRACT:

Background

Reverse phase protein arrays (RPPA) have been demonstrated to be a useful experimental platform for quantitative protein profiling in a high-throughput format. Target protein detection relies on the readout obtained from a single detection antibody. For this reason, antibody specificity is a key factor for RPPA. RNAi allows the specific knockdown of a target protein in complex samples and was therefore examined for its utility to assess antibody performance for RPPA applications.

Results

To proof the feasibility of our strategy, two different anti-EGFR antibodies were compared by RPPA. Both detected the knockdown of EGFR but at a different rate. Western blot data were used to identify the most reliable antibody. The RNAi approach was also used to characterize comm

SUBMITTER: Mannsperger HA 

PROVIDER: S-EPMC3022873 | biostudies-literature | 2010 Dec

REPOSITORIES: biostudies-literature