PP1 Catalytic Subunit Antibody (319319)

Detection of Human/Mouse/Rat PP1ca by Western Blot.

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, Nb2-11 rat lymphoma cell line, and Neuro-2A mouse neuroblastoma cell line. PVDF membrane was probed with 1 µg/mL of Human/Mouse/Rat PP1 Catalytic Subunit Monoclonal Antibody (Catalog # MAB3000) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Specific bands were detected for PP1 catalytic subunits at approximately 39 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of PP1 Catalytic Subunit by Western Blot

Akt and PP control C-peptide-induced beta -catenin phosphorylation. (A) The isolated human endometrial stromal cells (HESCs) were used as the undecidualized HESCs (UnD-ESCs) and decidualized HESCs (D-ESCs) on day 2 after the induction of decidualization in the presence of 8-Br-cAMP. UnD-ESCs and D-ESCs were serum-starved for 48 h and then incubated with 50 nM C-peptide for 10 min. The cells were lysed and subjected to western blot analysis. (B) UnD-ESCs and D-ESCs were lysed and GPR146 mRNA level was analyzed using qRT-PCR. (C) UnD-ESCs were pretreated with 1 μM Akti for 1 h and treated as described in (A). (D) UnD-ESCs were treated as described in (C) and protein phosphatase (PP) activity was measured. (E) UnD-ESCs were treated as described in (A) in pretreatment with 10 μM okadaic acid for 1 h. (F) After treatment with C-peptide for 10 min, UnD-ESCs were lysed and then immunoprecipitated with IgG or anti-beta -catenin. The PP2A levels of the immunoprecipitates were normalized using the beta -catenin level. (G) UnD-ESCs were transduced using short hairpin RNAs (shRNAs), selected for 5 days with 1.5 μM puromycin and treated as (A). (A,C,E,G) The intensities of western blots were quantified using ImageJ software. The relative phosphorylation level or protein expression level were calculated by comparing them to total protein or tubulin level, respectively. Asterisks (*) indicate significant differences (p < 0.05) when compared to C-peptide untreated control or C-peptide treated UnD-ESCs control. Akti, Akt inhibitor; OA, Okadaic acid; shScram, shRNA for Scramble; shPPP1C, shRNA for PPP1C. Image collected and cropped by CiteAb from the following open publication (https://www.frontiersin.org/articles/10.3389/fcell.2022.800181/full), licensed under a CC-BY license. Not internally tested by R&D Systems.