Spectrophotometric method for quantitative determination of nonionic, ionic and zwitterionic detergents.
Detergents serve as means of solubilizing biological membranes and thus play an important role in purification and characterization of membrane proteins. We report here a simple method to estimate the amount of detergent bound to a protein or present in an aqueous solution. The method is based on the turbidity caused by the addition of a detergent to triolein. Detergent bound to an integral membrane protein, lysophosphatidic acid acyltransferase, was separated by native gel electrophoresis and the amount of detergent bound to the same was estimated. This method is applicable for Triton X-100, sodium dodecyl sulfate and zwitterionic detergent, and was validated in the presence of reagents commonly used in membrane protein solubilization and purification.
Rajakumari
Sona
S
Lipid Laboratory, Department of Biochemistry, Indian Institute of Science, Bangalore 560012, India.
Srinivasan
Malathi
M
Rajasekharan
Ram
R
D000217:Acyltransferases / Q000737:chemistry / Q000302:isolation & purification; D010367:Arachis / Q000737:chemistry; D003902:Detergents / Q000032:analysis*; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification; D013053:Spectrophotometry / Q000379:methods
D003902:Detergents; D008565:Membrane Proteins; D000217:Acyltransferases