| First Author | Jackson TC | Year | 2015 |
| Journal | Sci Rep | Volume | 5 |
| Pages | 9377 | PubMed ID | 25827478 |
| Mgi Jnum | J:287126 | Mgi Id | MGI:6216954 |
| Doi | 10.1038/srep09377 | Citation | Jackson TC, et al. (2015) Detection of PHLPP1alpha/beta in human and mouse brain by different anti-PHLPP1 antibodies. Sci Rep 5:9377 |
| abstractText | 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 - PHLPP1alpha (~140-150 kDa) and PHLPP1beta (~180-190 kDa). Commercial antibodies are widely used to characterize PHLPP1 proteins in cells/tissues. Here we validate five different antibodies to detect PHLPP1alpha/beta by Western blot using PHLPP1 WT/KO mice. All antibodies recognize PHLPP1beta in brain. Only a single antibody (Cosmo Bio Co) detects PHLPP1alpha (~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 PHLPP1alpha 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. |
