产品介绍
Rab35 Pull-Down Activation Assay Kit
Cat. # 82801
Introduction
A. Background
The Rab family of GTPases regulates eukaryotic vesicular membrane traffic. Rab35 is with both plasma membrane and endosomal localization and has been implicated in diverse processes that include T-cell receptor recycling, oocyte yolk protein recycling and cytokinesis. Rab35 regulates neurite outgrowth in neuronal-like cells and can induce protrusions.
Currently there is no direct assay to measure the activation of Rab35 GTPases.
The Rab35 Activation Assay Kit is based on the configuration-specific monoclonal antibody that specifically recognizes Rab35-GTP, but not Rab35-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.
B. Assay Principle
The Rab35 Activation Assay Kit uses configuration-specific anti-Rab35-GTP Mouse monoclonal antibody to measure Rab35-GTP levels in cell extracts or in vitro GTPγS loading Rab35 activation assays. Anti-Rab35-GTP mouse monoclonal antibody is first incubated with cell lysates containing Rab35-GTP. Next, the GTP-bound Rab35 is pulled down by protein A/G agarose. Finally, the precipitated Rab35-GTP is detected through immunoblot analysis using Anti-Rab35 Rabbit Polyclonal Antibody.
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