Product Description

Arf-like protein 1 （Arl1） is a member of the Arf family of regulatory GTPases, within the Ras

superfamily of GTPases, and with highly conserved orthologs throughout eukaryotes. Arl1 is essential

for early embryonic development in Drosophila and in Caenorhabditis elegans. Arl1 is most similar in

primary sequence, cellular location, and function （regulation of membrane traffic） to Arf1–6 and even

shares several common binding partners. In addition to its function in membrane traffic at the

Golgi/trans-Golgi network, there are reports indicating a possible role for Arl1 in ion homeostasis in

yeast.

Currently there is no direct assay to measure the activation of Arl1 GTPases.

NewEast Biosciences Arl1 Activation Assay Kit is based on the configuration-specific monoclonal

antibody that specifically recognizes Arl1-GTP, but not Arl1-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-Arl1-GTP monoclonal antibodies can also be used to monitor the activation of Arl1 in cells

and in tissues by immunohistochemistry.

NewEast Biosciences Arl1 Activation Assay Kit provides a simple and fast method to monitor the

activation of Arl1. Each kit provides sufficient quantities to perform 20 assays.

Assay Principle

NewEast Biosciences bases on the configuration-specific anti-Arl1-GTP

monoclonal antibody to measure the active Arl1-GTP levels, either from cell extracts or from in vitro

GTPγS loading Arl1 activation assays. Briefly, anti-active Arl1 mouse monoclonal antibody will be

incubated with cell lysates containing Arl1-GTP. The bound active Arl1 will then be pulled down by

protein A/G agarose. The precipitated active Arl1 will be detected by immunoblot analysis using anti-

Arl1 rabbit polyclonal antibody or anti-Arl1 mouse monoclonal antibody.

Kit Components

1. Anti-active Arl1, Mouse Monoclonal Antibody （Catalog No. 26924）: One vial – 22 μL （1

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

specifically recognizes Arl1-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-Arl1, Mouse monoclonal Antibody （Catalog No. 26056）: 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.