Antibody was affinity purified using an epitope specific to ROCK1 immobilized on solid support. The epitope recognized by A300-457A maps to a region between residue 1300 and the C-terminus (residue 1354) of human Rho-associated, coiled-coil containing protein kinase 1 using the numbering given in Swiss-Prot entry Q13464 (GeneID 6093). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
p160 ROCK-1, P160ROCK, renal carcinoma antigen NY-REN-35, rho-associated protein kinase 1, Rho-associated, coiled-coil-containing protein kinase 1, Rho-associated, coiled-coil-containing protein kinase I, ROCK-I
between 1300 and C-term
2 - 8°C
Detection of human and mouse ROCK1 by western blot and immunoprecipitation. Samples: Whole cell lysate from HeLa (15 and 50 µg for WB; 1 mg for IP, 20% of IP loaded), HEK293T (T; 50 µg), Jurkat (J; 50 µg) and mouse NIH 3T3 (M; 50 µg) cells. Antibodies: Affinity purified rabbit anti-ROCK1 antibody A300-457A (lot A300-457A-2) used for WB at 0.04 µg/ml (A) and 1 µg/ml (B) and used for IP at 6 µg/mg lysate. ROCK1 was also immunoprecipitated by a previous lot (lot A300-457A-2) of this antibody. Detection: Chemiluminescence with exposure times of 10 seconds (A) and 1 second (B).