CST, 66256T, PPP1R14A/CPI-17 (F1I6X) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying PPP1r14a. Validated for WB,IP,IHC,IF. Available in 2 sizes. Highly specific and rigorously validated in-house, PPP1R14A/CPI-17 (F1I6X) Rabbit Monoclonal Antibody (CST #66256) is ready to ship. Product Usage Information Western Blotting: 1:1000 Immunoprecipitation: 1:200 Immunohistochemistry (Paraffin): 1:50 - 1:200 Immunofluorescence (Immunocytochemistry): 1:1000 - 1:4000 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. Protocol Available protocols: Western Blotting, Immunoprecipitation, Immunohistochemistry (Paraffin), Immunofluorescence (Immunocytochemistry) Specificity / Sensitivity PPP1R14A/CPI-17 (F1I6X) Rabbit Monoclonal Antibody recognizes endogenous levels of total PPP1R14A/CPI-17 protein. This antibody does not cross-react with PPP1R14B/PHI-1. This antibody detects a 70 kDa protein of unknown identity in some cell lines. Non-specific staining was observed at the apical membrane of normal human colon epithelium by immunohistochemistry. Immunocytochemistry reactivity is human only. Species Reactivity: Human, Mouse, Rat Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human PPP1R14A/CPI-17 protein. Background Protein kinase C-potentiated inhibitor protein of 17 kDa (CPI-17), encoded by the protein phosphatase 1 (PP1) regulatory subunit 14A ( ) gene, promotes smooth muscle contraction by inhibiting the activity of the PP1 holoenzyme, smooth muscle myosin phosphatase (MYPT1) (1). CPI-17 belongs to a family of proteins, including PPP1R14B/PHI-1, PPP1R14C/KEPI, and PPP1R14D/GEPI, that each specifically inhibits a subset of PP1 holoenzymes (1). CPI-17 expression, localization, and enzymatic activity are driven by a combination of epigenetic and posttranslational modifications, all of which have downstream effects on PP1-dependent cellular processes. CPI-17 is a substrate for PKC, ROCK1, ZIPK, MST3/MST4, and CaMKII, with established phosphorylation target sites at Ser12, Thr38, and Ser130 (1-6). Additional putative phosphorylation sites with as yet undefined upstream kinases may be regulated by luteinizing hormone signaling in rodent ovary (7). CPI-17 is translocated from the cytoplasm to the nucleus via the importin complex, and phosphorylation of CPI-17 at Ser12 disrupts the N-terminal nuclear localization signal (8). Nuclear CPI-17 inhibits PP1-mediated dephosphorylation of histone H3, and CPI-17 protein knockdown by siRNA decreased PANC-1 pancreatic carcinoma cell proliferation (8). Thr38 phosphorylation increases CPI-17 inhibitory activity 1,000-fold, and can be induced by upstream kinase agonists (9,10). In a pan-cancer analysis, PPP1R14A mRNA and protein expression were found to be decreased across a wide variety of cancer types, including breast, colon, and ovarian cancers, kidney renal clear cell carcinoma, lung adenocarcinoma, and uterine corpus endometrial carcinoma (11). The pan-cancer expression profile correlated with increased PPP1R14A promoter methylation (11). Increased gene silencing by DNA methylation was also observed in nearly 80% of non-Hodgkin lymphoma patient samples relative to control B lymphocytes (12). Alternate Names 17 kDa PKC-potentiated inhibitory protein of PP1; 17-KDa protein; 17-kDa PKC-potentiated inhibitory protein of PP1; 17-KDa protein; CPI-17; CPI17; PKC-potentiated inhibitory protein of PP1; PP14A; PPP1INL; PPP1R14A; Protein kinase C-potentiated inhibitor protein of 17 kDa; protein phosphatase 1 regulatory inhibitor subunit 14A; Protein phosphatase 1 regulatory subunit 14A; protein phosphatase 1, regulatory (inhibitor) subunit 14A Specification REACTIVITY: H M R SENSITIVITY: Endogenous MW (kDa): 19, 17 Source/Isotype: Rabbit IgG