81201 RheB Activation Assay Kit_详细资料

产品信息

Product Description
Small GTPases are a super-family of cellular signaling regulators. RheB is a member of the
Ras-superfamily GTPases. RheB has been shown to interact with C-Raf, Mammalian target of
rapamycin （mTOR）, TSC2, Ataxia angiectasia mutated （ATM）, KIAA1303 and Ataxia angiectasia
and Rad3 related.
Currently there is no direct assay to measure the activation of RheB GTPases.
NewEast Biosciences RheB Activation Assay Kit is based on the configuration-specific monoclonal
antibody that specifically recognizes RheB-GTP, but not RheB-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-RheB-GTP monoclonal antibodies can also be used to monitor the activation of RheB in cells
and in tissues by immunohistochemistry.
NewEast Biosciences RheB Activation Assay Kit provides a simple and fast method to monitor the
activation of RheB. Each kit provides sufficient quantities to perform 20 assays.
Assay Principle
NewEast Biosciences bases on the configuration-specific anti-RheB-GTP
monoclonal antibody to measure the active RheB-GTP levels, either from cell extracts or from in vitro
GTPγS loading RheB activation assays. Briefly, anti-active RheB mouse monoclonal antibody will be
incubated with cell lysates containing RheB-GTP. The bound active RheB will then be pulled down by
protein A/G agarose. The precipitated active RheB will be detected by immunoblot analysis using anti-
RheB rabbit polyclonal antibody.
Kit Components
1. Anti-active RheB, Mouse Monoclonal Antibody （Catalog No. 26910）: One vial – 22 μL （1
mg/ml） in PBS, pH 7.4, containing 50% glycerol and 0.05% sodium azide. This antibody
specifically recognizes RheB-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-RheB, Rabbit Polyclonal Antibody （Catalog No. 21098）: 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.

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