Configuration-specific Monoclonal Antibody Based

Ran Activation Assay Kit

Catalog Number：CR-W81701

20 assays

Product Description

Small GTPases are a super-family of cellular signaling regulators. Ran (Ras-related nuclear protein) is a

member of the Ras-superfamily GTPases. Ran is invovled in the transport of proteins across the nuclear

envelope, as well as in microtubule organization during mitosis.

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

Bioyears Biosciences Ran Activation Assay Kit is based on the configuration-specific monoclonal

antibody that specifically recognizes Ran-GTP, but not Ran 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 Ran GTP monoclonal antibodies can also be used to monitor the activation of Ran in cells

and in tissues by immunohistochemistry.

Bioyears Biosciences Ran Activation Assay Kit provides a simple and fast method to monitor the

activation of Ran. Each kit provides sufficient quantities to perform 20 assays.

Assay Principle

Bioyears Biosciences Ran Activation Assay Kit bases on the configuration-specific anti Ran GTP

monoclonal antibody to measure the active Ran GTP levels, either from cell extracts or from in vitro

GTPγS loading Ran activation assays. Briefly, anti active Ran mouse monoclonal antibody will be

incubated with cell lysates containing Ran GTP. The bound active Ran will then be pulled down by

protein A/G agarose. The precipitated active Ran will be detected by immunoblot analysis using anti Ran rabbit polyclonal antibody.

Kit Components

1. Anti active Ran, Mouse Monoclonal Antibody (Catalog No. CR-M26915): One vial – 22 µL (1

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

   specifically recognizes Ran 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. 30303): 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 Ran, Rabbit Polyclonal Antibody (Catalog No. CR-R21097): One vial – 22 µL (1 mg/ml) in

   PBS, pH 7.4, contained 50% glycerol.

5. 100 X GTPγS (Catalog No. 30302): 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.

Storage

Store all kit components at 4ºC until their expiration dates.