Chemical structure of Y-27632.
Western blot analysis of extracts from C2C12, 293, and NIH/3T3 cells, untreated (-) or treated with Y-27632 (10 μM, 1 hr; +), using Phospho-MYPT1 (Thr853) Antibody #4563 (upper) and MYPT1 Antibody #2634 (lower).
Y-27632 is supplied as a lyophilized powder. For a 10 mM stock, reconstitute the 1 mg in 312.2 µl DMSO. Working concentrations and length of treatment can vary depending on the desired effect, but it is typically used at 1-10 µM for 15 min-4 hr.
Store lyophilized or in solution at -20ºC, desiccated. In lyophilized form, the chemical is stable for 24 months. Once in solution, use within 3 months to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.
C14H21N3O · 2HCl
Y-27632 is a potent and selective ATP-competitive inhibitor of Rho-associated kinases (ROCK), with Ki values of 0.22 µM and 0.30 µM for ROCK1 and ROCK2, respectively. In vitro kinase assays demonstrate that Y-27632 exhibits a 20-fold greater preference for ROCK when compared to citron kinase and protein kinase N (1). Research studies show that Y-27632 treatment of cells leads to inhibition of ROCK phosphorylation of myosin phosphatase subunit 1 (MYPT1) at both Thr853 and Thr696, with a much greater inhibition of Thr853 phosphorylation (2,3). Y-27632 selectively inhibits smooth-muscle contraction by inhibiting Ca2+ sensitization (4). Additional research indicates that Y-27632 inhibits dissociation-induced apoptosis of cultured human embryonic stem cells over numerous passages and increases cloning efficiency (5).
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