Related Experiment Videos
Spectrophotometric method for quantitative determination of nonionic, ionic and zwitterionic detergents.
Sona Rajakumari1, Malathi Srinivasan, Ram Rajasekharan
1Lipid Laboratory, Department of Biochemistry, Indian Institute of Science, Bangalore 560012, India.
Journal of Biochemical and Biophysical Methods
|June 8, 2006
Summary
A new turbidity-based method accurately quantifies detergent amounts in protein solutions and bound to membrane proteins. This technique aids in detergent solubilization and purification processes.
Area of Science:
- Biochemistry
- Protein Chemistry
- Membrane Biology
Background:
- Detergents are crucial for solubilizing biological membranes.
- Accurate detergent quantification is vital for membrane protein purification and characterization.
Purpose of the Study:
- To develop a simple and versatile method for estimating detergent amounts.
- To quantify detergent bound to membrane proteins.
Main Methods:
- A turbidity-based assay using triolein was employed.
- Native gel electrophoresis was used to separate detergent-bound proteins.
- The method was validated with common detergents and reagents.
Main Results:
- The method successfully estimated detergent bound to lysophosphatidic acid acyltransferase.
- Applicable to Triton X-100, sodium dodecyl sulfate, and zwitterionic detergents.
- Validated in the presence of common membrane protein solubilization reagents.
Conclusions:
- This turbidity assay provides a simple and reliable way to measure detergent.
- It is applicable to various detergents and experimental conditions relevant to membrane protein studies.