Antibody was affinity purified using an epitope specific to ROCK2 immobilized on solid support. The epitope recognized by A300-047A maps to a region between residues 1320 and the C-terminus (residues 1388) of human ROCK2 using the number given in entry NP_004841.1 (GeneID 9475) Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
p164 ROCK-2, Rho kinase 2, rho-associated protein kinase 2, Rho-associated, coiled-coil-containing protein kinase 2, rho-associated, coiled-coil-containing protein kinase II, ROCK-II
between 1320 and C-term
2 - 8°C
Detection of human ROCK2 by western blot. Samples: Whole cell lysate (50 µg) from HeLa and HEK293T cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-ROCK2 antibody A300-047A (lot A300-047A-1) used for WB at 0.04 µg/ml. Detection: Chemiluminescence with an exposure time of 10 seconds.
Detection of human ROCK2 by western blot and immunoprecipitation. Samples: Whole cell lysate (WCL) from HeLa cells. In A, ROCK2 from 5, 10 and 20 µg of WCL or immunoprecipitated ROCK2 was detected. In B, ROCK2 was immunoprecipitated from 1 mg of WCL. Antibodies: A) Rabbit anti-ROCK2 antibody A300-047A was used at 0.06 µg/ml to detect ROCK2 in WCL and ROCK2 immunoprecipitated using an antibody generated to a recombinant fragment. B) Rabbit anti-ROCK2 antibodies BL963 and A300-047A were used at 5 µg/mg lysate to immunoprecipitate ROCK2, which was detected by western blot using an antibody generated to a recombinant fragment of ROCK2. Detection: Chemiluminescence with exposure times of 5 to 15 seconds.