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Testing Data

ROCK Activity Assay Kit

96-Well ROCK Activity Assay Kit

Synonyms
ROCK Activity; 96-Well ROCK Activity Assay Kit; ROCK Activity assay kit
Ordering
For Research Use Only!
Preparation and Storage
Store active ROCK-II at -80 degree C, ATP Solution at -20 degree C and all other kit components at 4 degree C until their expiration dates. Avoid multiple freeze/thaw cycles.

Testing Data

Testing Data

Testing Data #2

Testing Data #2

Testing Data #3

Testing Data #3
Related Product Information for ROCK Activity 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.
Product Categories/Family for ROCK Activity assay kit
References
1. Etienne-Manneville, S., and Hall, A. (2002) Nature 420, 629-635.
2. Riento, K., and Ridley, A. J. (2003) Nat. Rev. Mol. Cell. Biol. 4, 446-456.
3. Leung, T., Manser, E., Tan, L., and Lim, L. (1995) J. Biol. Chem. 270, 29051-29054.
4. Matsui, T., Amano, M., Yamamoto, T., Chihara, K., Nakafuku, M., Ito, M., Nakano, T., Okawa, K., Iwamatsu, A., and Kaibuchi, K. (1996) EMBO J. 15, 2208-2216.
5. Totsukawa, G, et al., J. Cell Biol. (1999) 144, 735-744.
1. Weber, M et al. (2014). MiRNA-155 targets myosin light chain kinase and modulates actin cytoskeleton organization in endothelial cells. Am. J. Physiol. Heart Circ. Physiol. 306:H1192-H1203.
2. Narimatsu, T. et al. (2013). Disruption of Cell-Cell Junctions and Induction of Pathological Cytokines in the Retinal Pigment Epithelium of Light-Exposed Mice. Invest. Opthalmol. Vis. Sci. 54:4555-4562.
3. Singh, J. et al. (2013). Role of PKC and RhoA/ROCK Pathways in the Spontaneous Phasic Activity in the Rectal Smooth Muscle. Am J Physiol Gastrointest Liver Physiol. 304:G723-G731.
4. Barcia, C. et al. (2012). ROCK/Cdc42-Mediated Microglial Motility and Gliapse Formation Lead to Phagocytosis of Degenerating Dopaminergic Neurons in vivo. Sci Rep. 2:809.
5. Zhang, W. et al. (2012). The Small GTPase RhoA Regulates the Contraction of Smooth Muscle Tissues by Catalyzing the Assembly of Cytoskeletal Signaling Complexes at Membrane Adhesion Sites. J. Biol. Chem. 287: 33966-34008.
6. Burger, D. et al. (2012). Microparticles Induce Cell Cycle Arrest Through Redox-Sensitive Processes in Endothelial Cells: Implications in Vascular Senescence. JAHA. 1: e001842.
7. DiPaolo, B.C. et al. (2012). Rho Kinase Signaling Pathways During Stretch in Primary Alveolar Epithelia. Am J Lung Cell Mol Physiol. 302: L992-L1002.
8. Burger, D. et al. (2011). Endothelial Microparticle Formation by Angiotensin II is Mediated via Ang II Receptor Type I/NADPH Oxidase/Rho Kinase Pathways Targeted to Lipid Rafts. Arterioscler Thromb Vasc Biol. 31:1898-1907.
9. Yotova, I.Y. et al. (2011). Abnormal Activation of Ras/Raf/MAPK and RhoA/ROCKII Signaling Pathways in Eutopic Endometrial Stromal cells of Patients with Endometriosis. Hum. Reprod. 10.1093/humrep/der010.
10. Haas, B. et al. (2009). Protein Kinase G Controls Brown Fat Cell Differentiation and Mitochondrial Biogenesis. Sci. Signal. 2:ra78.

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Product Notes

The ROCK Activity (Catalog #AAA168354) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "ROCK Activity, Assay Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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