ROCK1 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Rho-associated kinase I (ROCK1, ROK beta, p160ROCK) is a serine-threonine protein kinase and an effector of the small GTPase Rho. With N-terminus protein kinase domain and C-terminus Rho-binding domain/pleckstrin homology domain, ROCK1 can be activated by either RhoA or RhoB. In Rho specific ROCK1 activation, a Rho protein binds to the Rho-binding domain and induces a conformational change which opens the kinase domain for the phosphorylation of downstream effectors (1). Activated ROCK1 phosphorylates various signaling proteins, such as myosin light chain phosphatase, LIM kinases, and ezrin-radixin-moesin proteins. Caspase-3 also activates ROCK1; Caspase-3 cleaves ROCK1 at DETD1113/G sequence and removes its inhibitory c-terminal domain. This activation, independent of Rho activity, leads to apoptotic membrane blebbing (2). ROCK1 is involved in regulating actin cytoskeleton assembly, cell migration, centromere positing, smooth muscle contraction, and neurite outgrowth. Its involvement in tumor invasion, hypertension, and bronchial asthma makes ROCK1 an ideal target for drug development (3).

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBP2-07756

• ROCK1 Antibodies
• ROCK1 Lysate
• ROCK1 Proteins
• ROCK1 Proteins and Enzyme