Product Highlight

• Configuration-specific monoclonal antibody that recognizes Rap1-GTP, but not Rap1-GDP
• The only available Rap1 activation assay kit directly measures the level of Rap1-GTP
• Providing shorter experimental time
• Could be used to monitor the activation of Rap1 in cells and in tissues by immunohistochemistry

Product Description

Small GTPases are a super-family of cellular signaling regulators. The small Ras-like GTPase Rap1 is an evolutionary conserved protein that originally gained interest because of its capacity to revert the morphological phenotype of Ras-transformed fibroblasts. Rap1 is regulated by a large number of stimuli that include growth factors and cytokines, but also physical force and osmotic stress. Rap1 was shown to regulate multiple basic cellular processes. The best studied aspect of Rap1 function in endothelial cells involved its role in regulation of cell-cell junction formation and remodeling.

Currently there is no direct assay to measure the activation of Rap1 GTPases.

NewEast Biosciences Rap1 Activation Assay Kit is based on the configuration-specific monoclonal antibody that specifically recognizes Rap1-GTP, but not Rap1-GDP. Given the high affinity of monoclonal antibodies to their antigens, the activation assay could be performed in a short time. This assay provides the reliable results with consistent reproducibility.

These anti-Rap1-GTP monoclonal antibodies can also be used to monitor the activation of Rap1 in cells and in tissues by immunohistochemistry.

NewEast Biosciences Rap1 Activation Assay Kit provides a simple and fast method to monitor the activation of Rap1. Each kit provides sufficient quantities to perform 20 assays.

 

Assay Principle

NewEast Biosciences Rap1 Activation Assay Kit bases on the configuration-specific anti-Rap1-GTP monoclonal antibody to measure the active Rap1-GTP levels, either from cell extracts or from in vitro GTPΓS loading Rap1 activation assays. Briefly, anti-active Rap1 mouse monoclonal antibody will be incubated with cell lysates containing Rap1-GTP. The bound active Rap1 will then be pulled down by protein A/G agarose. The precipitated active Rap1 will be detected by immunoblot analysis using anti- Rap1 rabbit polyclonal antibody.

Publications:

1.  Rap1 GTPase Activation and Barrier Enhancement in RPE Inhibits Choroidal Neovascularization In Vivo     PLoS One. 2013 Sep 10;8(9):e73070