ATPase/GTPase Assay Kit

Catalog No. ABIN1000250

Product Details

Target: ATPase/GTPase

Application: Activity Assay (AcA)

Characteristics: High sensitivity: detection of as little of 2 pmoles of free phosphate.
Fast and convenient: single reagent, homogeneous mix-and-measure assay allows quantitation of enzyme activity within 30 minutes.
Robust and amenable to HTS: detection at 620nm greatly reduces potential interference by colored compounds. Z' factors of >0.7 are observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems.

Components: Reagent: 50 mL. Assay Buffer: 10 mL. Standard: 1mL 1 mM phosphate.

Application Details

Application Notes: Determination of ATPase and GTPase activity.
Drug Discovery: high-throughput screen for ATPase or GTPase inhibitors.

Comment:

Use ultrapure ATP and GTP. The provided 2x assay buffer contains 40 mM Tris, 80 mM NaCl, 8 mM MgAc2, 1 mM EDTA, pH 7.5. Other buffers (Hepes, Mes, Mops) can be used. Assay is compatible with 1 mM DTT, 2mM -mercaptoethanol, 0.5 mg/mL BSA and 5% DMSO.

Restrictions: For Research Use only

Handling

Storage: 4 °C

References

Sheikha, Al-Shaer, Taha: "Some sulfonamide drugs inhibit ATPase activity of heat shock protein 90: investigation by docking simulation and experimental validation." in: Journal of enzyme inhibition and medicinal chemistry, Vol. 26, Issue 5, pp. 603-9, (2011) (PubMed).

Hussey, Chaudhury, Dawson, Lindner, Knudsen, Wilce, Merrick, Howe: "Identification of an mRNP complex regulating tumorigenesis at the translational elongation step." in: Molecular cell, Vol. 41, Issue 4, pp. 419-31, (2011) (PubMed).

Binder, Eberle, Seitz, Mücke, Hüber, Kiani, Kaderali, Lohmann, Dalpke, Bartenschlager: "Molecular mechanism of signal perception and integration by the innate immune sensor retinoic acid-inducible gene-I (RIG-I)." in: The Journal of biological chemistry, Vol. 286, Issue 31, pp. 27278-87, (2011) (PubMed).

Bertol, Rigotto, de Pádua, Kreis, Barardi, Braga, Simões: "Antiherpes activity of glucoevatromonoside, a cardenolide isolated from a Brazilian cultivar of Digitalis lanata." in: Antiviral research, Vol. 92, Issue 1, pp. 73-80, (2011) (PubMed).

Suematsu, Yokobori, Morita, Yoshinari, Ueda, Kita, Takeuchi, Watanabe: "A bacterial elongation factor G homologue exclusively functions in ribosome recycling in the spirochaete Borrelia burgdorferi." in: Molecular microbiology, Vol. 75, Issue 6, pp. 1445-54, (2010) (PubMed).

Hu, Lang, Krebsbach: "Digoxigenin modification of adenovirus to spatially control gene delivery from chitosan surfaces." in: Journal of controlled release : official journal of the Controlled Release Society, Vol. 135, Issue 3, pp. 250-8, (2009) (PubMed).

Target Details

Target: ATPase/GTPase

Background: Quantitative determination of ATPase or GTPase activity by colorimetric (620nm) method. High-throµghput screening for ATPase/GTPase inhibitors.
Procedure: 30 min.

ATPases and GTPases catalyze the decomposition of ATP or GTP into ADP or GDP and free phosphate ion. These enzymes play key roles in transport, signal transduction, protein biosynthesis and cell differentiation. The ATPase/GTPase Assay Kit offers a highly sensitive method for determining ATPase/GTPase activities in a microplate format. Its proprietary formulation features a single reagent for accurate determination of enzyme activity in 30 min at room temperature. The improved malachite green reagent forms a stable dark green color with liberated phosphate, which is measured on a plate reader (600 - 660 nm).