Small GTPases are a super-family of cellular signaling regulators. Rab11 has been shown to control traffic through the recycling endosome. Currently there is no direct assay to measure the activation of Rab11 GTPases.

    Bioyears Biosciences Rab11 Activation Assay Kit is based on the configuration-specific monoclonal antibody that specifically recognizes Rab11 GTP, but not Rab11-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 Rab11 GTP monoclonal antibodies can also be used to monitor the activation of Rab11 in cells and in tissues by immunohistochemistry. Bioyears Biosciences Rab11 Activation Assay Kit provides a simple and fast method to monitor theactivation of Rab11. Each kit provides sufficient quantities to perform 20 assays.

Bioyears Biosciences Rab11 Activation Assay Kit bases on the configuration-specific anti Rab11 GTP monoclonal antibody to measure the active Rab11 GTP levels, either from cell extracts or from in vitro GTPγS loading Rab11 activation assays. Briefly, anti active Rab11 mouse monoclonal antibody will be incubated with cell lysates containing Rab11 GTP. The bound active Rab11 will then be pulled down by protein A/G agarose. The precipitated active Rab11 will be detected by immunoblot analysis using an anti Rab11 rabbit polyclonal antibody. 

1. Anti active Rab11, Mouse Monoclonal Antibody (Catalog No. 26919): One vial – 22 µL (1mg/ml) in PBS, pH 7.4, containing 50%glycerol and 0.05% sodium azide. This antibody specifically recognizes Rab11 GTP from all vertebrates.

2. Protein A/G Agarose (Catalog No. 30301): One vial – 400 µL of 50% slurry.

3. 5X Assay/Lysis Buffer (Catalog No. 30302): One bottle – 30 mL of 250 mM Tris-HCl, pH 8, 750mM NaCl, 50 mM MgCl2, 5 mM EDTA,5% Triton X-100.

4. Anti Rab11, Rabbit Polyclonal Antibody (Catalog No. 21157): One vial – 22 µL (1 mg/ml)in PBS, pH 7.4, contained 50% glycerol.

5. 100 X GTPγS (Catalog No. 30303): One vial –100 µl at 10 mM, use 5 µL of GTPγS for GTP-labeling of 0.5 mL of cell lysate.

6. 100 X GDP (Catalog No. 30304): One vial –100 µl at 100 mM, use 5 µL of GDP forGDP-labeling of 0.5 mL of cell lysate.