Small GTPases are a super-family of cellular signaling regulators. The small Ras-like GTPase Rap1 is anevolutionary 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.

Bioyears 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 cellsand in tissues by immunohistochemistry.

Bioyears 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. 

1. Anti-active Rap1, Mouse Monoclonal Antibody (Catalog No. 26912): One vial – 22 µL (1

    mg/ml) in PBS, pH 7.4, containing 50% glycerol and 0.05% sodium azide. This antibody 

    specifically recognizes Rap1 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, 750 mM NaCl, 50 mM MgCl2, 5 mM EDTA, 5% Triton X-100.

4. Anti-Rap1, Rabbit Polyclonal Antibody (Catalog No. 21104): One vial – 100 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 for

    GDP-labeling of 0.5 mL of cell lysate.