Antibody was affinity purified using an epitope specific to PNUTS immobilized on solid support. The epitope recognized by A300-439A maps to a region between residue 900 and the C-terminus (residue 940) of human Phosphatase Nuclear Targeting Subunit (a.k.a. Protein Phosphatase 1, regulatory subunit 10) using the numbering given in TrEMBL entry Q96QC0 (GeneID 5514). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
CAT53, FB19, HLA-C associated transcript 53, MHC class I region proline-rich protein CAT53, p99, phosphatase 1 nuclear targeting subunit, PNUTS, PP1-binding protein of 114 kDa, PP1R10, Protein FB19, protein phosphatase 1, regulatory (inhibitor) subunit 10
between 900 and C-term
2 - 8°C
Detection of human PNUTS by western blot. Samples: Whole cell lysate (50 µg) from HeLa and HEK293T cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-PNUTS antibody A300-440A (lot A300-440A-1) used for WB at 0.2 µg/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.
Detection of human PNUTS by western blot and immunoprecipitation. Samples: Nuclear extract (NE; 15 and 45 µg for WB; 100 µg for IP) from HeLa cells. Antibodies: Affinity purified rabbit anti-PNUTS antibody A300-440A (lot A300-440A-1) used at 1 and 0.2 µg/ml for WB (A) and at 1 µg/100 µg NE for IP (B). immunoprecipitation of PNUTS was also performed with rabbit anti-PNUTS antibodies BL1929, BL1930 and A300-439A. immunoprecipitatesd PNUTS was blotted using BL1932 at 0.2 µg/ml. Detection: Chemiluminescence with exposure times of 10 seconds (A) and 3 seconds (B).