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Technical Data Sheet

RM2741

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ROCK-I (C-terminal), cleavage-specific
Mouse Monoclonal IgG1
Price
Size
Species Reactivity
MW

$295
200 μl
Hu, Rt, Ms
31 kDa

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Western blot analysis of human Jurkat cells untreated (lanes 1 & 3) or treated with camptothecin (lanes 2 & 4). The blots were probed with anti-ROCK-I (RM2721) (lanes 1 & 2) and anti-ROCK-I (C-terminal) cleavage-specific (RM2741) (lanes 3 & 4).

Application
Dilution


ELISA
1:500


WB
1:250



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
Rho-associated kinases (ROCKs) are a family of serine/threonine kinases that have important roles in Rho-mediated signaling. The structure of ROCKs (ROCK-I/ROKβ and ROCK-II/ROKα) includes an N-terminal catalytic domain, a coiled-coil region, and a C-terminal pleckstrin homology domain. The C-terminal region negatively regulates kinase activity, and this inhibitory regulation is controlled through C-terminal cleavage. Caspase-3-induced cleavage of ROCK-I and direct cleavage of ROCK-II by granzyme B activate ROCK. This leads to inhibition of myosin phosphatase and phosphorylation of myosin light chain, which causes myosin II-mediated changes in cytoskeletal dynamics. This pathway has been implicated as a downstream effector of Rho-mediated changes in cytokinesis, cell motility, apoptosis and muscle contraction.


Background References
Nakagawa, O. et al. (1996) FEBS Lett. 392:189.
Lee, J.H. et al. (2004) J. Cell. Biol. 167:327.
Sebbagh, M. et al. (2005) J. Exp. Med. 201:465.
Immunogen
Clone (154C1465) was generated from a.a. 1114-1126 adjacent to the caspase cleavage site in human ROCK-I. This sequence has high homology to rat and mouse ROCK-I, and has significant homology to the conserved region in ROCK-II.
Buffer and Storage
Mouse monoclonal purified with protein G chromatography is supplied in 200μl phosphate-buffered saline with 0.2% gelatin and 0.05% sodium azide. Store at 4°C, stable for 6 months. For long term storage, aliquot and store at -20°C.
Specificity
The antibody detects a 31 kDa* band corresponding to the C-terminal cleavage fragment of ROCK-I on SDS-PAGE immunoblots of human Jurkat cells treated with camptothecin.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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