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 Small GTPases are a super-family of cellular signaling regulators. Sar1 is a member of these smallGTPases. Sar1 functions as a molecular switch to control protein–protein and protein–lipid

interactions that direct vesicle budding from the ER. Sar1 is invloved in membrane trafficking, andregulates the assembly and disassembly of COPII coats.Currently there is no direct assay to measure the activation of Sar1 GTPases.

  Bioyears Biosciences Sar1 Activation Assay Kit is based on the configuration-specific monoclonalantibody that specifically recognizes Sar1 GTP, but not Sar1 GDP. Given the high affinity ofmonoclonal 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 Sar1 GTP monoclonal antibodies can also be used to monitor the activation of Sar1 in cellsand in tissues by immunohistochemistry.

    Bioyears Biosciences Sar1 Activation Assay Kit provides a simple and fast method to monitor the activation of Sar1. Each kit provides sufficient quantities to perform 20 assays.  

Bioyears Biosciences Sar1 Activation Assay Kit bases on the configuration-specific anti Sar1 GTP monoclonal antibody to measure the active Sar1 GTP levels, either from cell extracts or from in vitro GTPγS loading Sar1 activation assays. Briefly, anti active Sar1 mouse monoclonal antibody will be incubated with cell lysates containing Sar1 GTP. The bound active Sar1 will then be pulled down by protein A/G agarose. The precipitated active Sar1 will be detected by immunoblot analysis using anti Sar1 rabbit polyclonal antibody. 

1. Anti active Sar1, Mouse Monoclonal Antibody (Catalog No. 26916): One vial – 22 μL

    (1mg/ml) in PBS, pH 7.4, containing 50% glycerol and 0.05% sodium azide. This antibody

    specifically recognizes Sar1 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 50 mM HEPES (pH8),

    0.1mg/ml BSA ,0.1%Triton-100, 1 mM DTT.

4. Anti Sar1, Rabbit Polyclonal Antibody (Catalog No. 21090): 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 forGTP-

    labelingof 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.  

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