Tags

Type your tag names separated by a space and hit enter

Optimization of a malachite green assay for detection of ATP hydrolysis by solubilized membrane proteins.
Anal Biochem. 2012 Jul 15; 426(2):103-5.AB

Abstract

We studied the activity of the fluorescently labeled membrane transporter MalGFK(2), which transports maltose at the expense of ATP hydrolysis. We used a commercially available malachite green assay (SensoLyte MG phosphate assay kit; Anaspec) to quantify the liberated phosphate upon ATP hydrolysis. However, strong variations in phosphate concentration were measured when using the supplier's handling protocol. We optimized the protocol, taking into account the effects mediated by glycerol, SDS, and fluorescent label on the sample. As a result we obtained highly reproducible phosphate concentration values under conditions optimal for solubilized membrane proteins.

Authors+Show Affiliations

Institut für Experimentalphysik, Freie Universität Berlin, D-14195 Berlin, Germany.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22507377

Citation

Repen, Boris, et al. "Optimization of a Malachite Green Assay for Detection of ATP Hydrolysis By Solubilized Membrane Proteins." Analytical Biochemistry, vol. 426, no. 2, 2012, pp. 103-5.
Repen B, Schneider E, Alexiev U. Optimization of a malachite green assay for detection of ATP hydrolysis by solubilized membrane proteins. Anal Biochem. 2012;426(2):103-5.
Repen, B., Schneider, E., & Alexiev, U. (2012). Optimization of a malachite green assay for detection of ATP hydrolysis by solubilized membrane proteins. Analytical Biochemistry, 426(2), 103-5. https://doi.org/10.1016/j.ab.2012.04.016
Repen B, Schneider E, Alexiev U. Optimization of a Malachite Green Assay for Detection of ATP Hydrolysis By Solubilized Membrane Proteins. Anal Biochem. 2012 Jul 15;426(2):103-5. PubMed PMID: 22507377.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Optimization of a malachite green assay for detection of ATP hydrolysis by solubilized membrane proteins. AU - Repen,Boris, AU - Schneider,Erwin, AU - Alexiev,Ulrike, Y1 - 2012/04/13/ PY - 2012/01/16/received PY - 2012/04/08/revised PY - 2012/04/10/accepted PY - 2012/4/18/entrez PY - 2012/4/18/pubmed PY - 2012/10/2/medline SP - 103 EP - 5 JF - Analytical biochemistry JO - Anal. Biochem. VL - 426 IS - 2 N2 - We studied the activity of the fluorescently labeled membrane transporter MalGFK(2), which transports maltose at the expense of ATP hydrolysis. We used a commercially available malachite green assay (SensoLyte MG phosphate assay kit; Anaspec) to quantify the liberated phosphate upon ATP hydrolysis. However, strong variations in phosphate concentration were measured when using the supplier's handling protocol. We optimized the protocol, taking into account the effects mediated by glycerol, SDS, and fluorescent label on the sample. As a result we obtained highly reproducible phosphate concentration values under conditions optimal for solubilized membrane proteins. SN - 1096-0309 UR - https://www.unboundmedicine.com/medline/citation/22507377/Optimization_of_a_malachite_green_assay_for_detection_of_ATP_hydrolysis_by_solubilized_membrane_proteins_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-2697(12)00228-X DB - PRIME DP - Unbound Medicine ER -