Detection of PHLPP1?/? in human and mouse brain by different anti-PHLPP1 antibodies.

ABSTRACT: Pleckstrin homology domain and leucine rich repeat protein phosphatase 1 (PHLPP1) is a member of the serine/threonine family of phosphatases. It has been studied in organs including brain, heart, pancreas, adipose, breast, and prostate. Human PHLPP1 encodes two splice variants - PHLPP1? (~140-150?kDa) and PHLPP1? (~180-190?kDa). Commercial antibodies are widely used to characterize PHLPP1 proteins in cells/tissues. Here we validate five different antibodies to detect PHLPP1?/? by Western blot using PHLPP1 WT/KO mice. All antibodies recognize PHLPP1? in brain. Only a single antibody (Cosmo Bio Co) detects PHLPP1? (~145-150?kDa). The other four antibodies detect a non-specific signal at ~150?kDa as evidenced by its abundance in PHLPP1 KO tissues. Results suggest Cosmo antibody is a better reagent to detect PHLPP1? by Western blot. In contrast, we found it unsuitable for immunofluorescence applications in brain. Our findings caution interpretation of the ~150?kDa band detected by some PHLPP1 antibodies in rodent and human tissues. Results also recapitulate the importance of including molecular weight standards in Western blot data to simplify retrospective analysis.

SUBMITTER: Jackson TC 

PROVIDER: S-EPMC4649889 | biostudies-literature | 2015 Apr

REPOSITORIES: biostudies-literature