ROCK Activity (ROCK) Assay Kit

Background: Members of the Rho family are essential regulatory components of the signaling pathway that direct cell motility, adhesion, and cytokinesis through reorganization of actin cytoskeleton. Rho is activated by extracellular signals such as lysophosphatidic acid (LPA). The actions of Rho are mediated by downstream Rho effectors. One of these effectors is Rho-associated kinase (ROCK). Two ROCK isoforms have been identified: ROCK-I (also known as ROKbeta) and ROCK-II (also known as Rho Kinase and ROKalpha). ROCK mediates Rho signaling and reorganizes actin cytoskeleton through phosphorylation of several substrates that contribute to the assembly of actin filaments and contractility. For example, ROCK inactivates myosin phosphatase through the specific phosphorylation of myosin phosphatase target subunit 1 (MYPT1) at Thr696, which results in an increase in the phosphorylated content of the 20-kDa myosin light chain (MLC20). 96-well ROCK Activity Assay Kit is an enzyme immunoassay developed for detection of the specific phosphorylation of MYPT1 at Thr696 by ROCK. A strip well microtiter plate is precoated with a recombinant MYPT1. After incubating the substrate wells with ROCK samples (such as purified kinase, cell or tissue lysate) the phosphorylated MYPT1 is detected by an anti-phospho-MYPT1 (Thr696) antibody (Figure 1). 96-well ROCK Activity Assay Kit provides a non-isotopic, sensitive and specific method to monitor ROCK activity using its physiological substrate; it can also be used in screening ROCK inhibitors. The kit has detection sensitivity limit of 200 pg of active ROCK-II. A recombinant active ROCK-II is also provided as a positive control. Each kit provides sufficient quantities to perform up to 96 assays.

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Small volumes of ROCK Activity assay kit vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

MBS168354 is an assay kit targeting ROCK Activity. Also known as 96-Well ROCK Activity Assay Kit.