Product Highlight

• Configuration-specific monoclonal anti-Rac-GTP antibody based
• This monoclonal antibody specifically recognizes Rac-GTP, but not Rac-GDP
• Providing shorter experimental time
• Providing reliable results with consistent reproducibility than other Rac activation assay kits
• Could be used to monitor the activation of Rac in cells and in tissues by immunohistochemistry

Product Description

Small GTPases are a super-family of cellular signaling regulators. Rac belongs to the Rho sub-family of GTPases that regulate cell motility, cell division, and gene transcription. GTP binding increases the activity of Rac, and the hydrolysis of GTP to GDP renders it inactive.

Currently the activation of Rac proteins is assayed with the binding of GTP-bound Rac to the p21-binding domain (PBD) of p21-activated protein kinase (PAK). This method is based on the observation that the active, GTP-bound Rac could bind to the PBD of PAK. However, the reproducibility of this method is poor. This is partially due to the relatively quick hydrolysis of GTP to GDP during the assay procedure, and the low binding affinity of PBD to Rac-GTP.

These anti-Rac-GTP monoclonal antibody can also be used to monitor the activation of Rac in cells and in tissues by immunohistochemistry.

NewEast Biosciences Rac Activation Assay Kit provides a simple and fast method to monitor the activation of Rac. Each kit provides sufficient quantities to perform 20 assays.

 

Assay Principle

NewEast Biosciences Rac Activation Assay Kit bases on the configuration-specific anti-Rac-GTP monoclonal antibody to measure the active Rac-GTP levels, either from cell extracts or from in vitro GTPγS loading Rac activation assays. Briefly, anti-active Rac mouse monoclonal antibody will be incubated with cell lysates containing Rac-GTP. The bound active Rac will then be pulled down by protein A/G agarose. The precipitated active Rac will be detected by immunoblot analysis using anti-Rac rabbit polyclonal antibody.

Publications:

1.  Phosphorylation of α-tubulin by protein kinase C stimulates microtubule dynamics in human breast cells Cytoskeleton Volume 71, Issue 4, pages 257–272, April 2014

2.  Immunofluorescence and Confocal Microscopy of Neutrophils Neutrophil Methods and Protocols Methods in Molecular Biology Volume 1124, 2014, pp 251-268

3.  A link between the nuclear-localized srGAP3 and the SWI/SNF chromatin remodeler Brg1 Molecular and Cellular Neuroscience Volume 60, May 2014, Pages 10–25

4.  SHP2 Phosphatase Promotes Mast Cell Chemotaxis toward Stem Cell Factor via Enhancing Activation of the Lyn/Vav/Rac Signaling Axis The Journal of Immunology April 14, 2014 1301155

5.  ROBO1, a tumor suppressor and critical molecular barrier for localized tumor cells to acquire invasive phenotype: Study in African-American and Caucasian prostate cancer models International Journal of Cancer Volume 135, Issue 11, pages 2493–2506, 1 December 2014

6.  Transcription factors NRF2 and nf-kb are coordinated effectors of the RHO family, GTP binding protein rac1 during inflammation J Biol Chem. 2014 May 30;289(22):15244-58

7.  Canonical and Non-canonical G-protein Signaling Helps Coordinate Actin Dynamics to Promote Macrophage Phagocytosis of Zymosan Mol Cell Biol. 2014 Nov 15;34(22):4186-99

8.  p53 Down Regulates PDGF-Induced Formation of Circular Dorsal Ruffles in Rat Aortic Smooth Muscle Cells PLoS One. 2014 Sep 23;9(9):e108257

9.  LPA-mediated migration of ovarian cancer cells involves translocalization of Gαi2 to invadopodia and association with Src and β-pix Cancer Lett. 2015 Jan 28;356(2 Pt B):382-91

10.  cAMP controls the restoration of endothelial barrier function after thrombin-induced hyperpermeability via Rac1 activation Physiological ReportsPublished 24 October 2014Vol. 2no. e12175DOI: 10.14814/phy2.12175

11.  Cisplatin at Sub-toxic Levels Mediates Integrin Switch in Lung Cancer Cells Anticancer Research December 2014 vol. 34 no. 12 7111-7117

12.  Semaphorin-3F suppresses the stemness of colorectal cancer cells by inactivating Rac1 Cancer letters. March 1, 2015Volume 358, Issue 1, Pages 76–84.

13.  Geometry sensing through POR1 regulates Rac1 activity controlling early osteoblast differentiation in response to nanofiber diameter Integr Biol (Camb). 2014 Dec 24

14.  Inhibition of Rac1 Activity in the Hippocampus Impairs the Forgetting of Contextual Fear Memory Mol Neurobiol. 2015 Jan 24