A novel method for detergent concentration determination.
A fast and precise method for detergent concentration determination is presented. (Patent applications for the method described here have been submitted (EP05011904 and US60/702,261). Depending on the interest of the scientific community, the system will be commercialized. (For further information contact Hervé-W. Rémigy at the e-mail address below.) A small droplet of the detergent solution is deposited on a piece of Parafilm M and side views are recorded by two orthogonally arranged TV cameras. The droplet contours are then approximated by ellipses to determine the contact angles. Comparison of the observed contact angle values to calibrated standard curves of known detergent concentrations gives the concentration of the detergent assessed. A range of commonly used detergents was studied to demonstrate the reproducibility and precision of this simple method. As a first application, the detergent binding capacity of the Escherichia coli galactose/proton symporter (GalP) was assessed. Aggregation of GalP was observed when <260 +/- 5 dodecyl-beta,D-maltoside molecules were bound to one GalP molecule. These measurements document the efficacy of the drop-shape based detergent concentration determination described.
Kaufmann
Thomas C
TC
M. E. Müller Institute for Microscopy at the Biozentrum, University of Basel, Basel, Switzerland.
Engel
Andreas
A
Rémigy
Hervé-W
HW
D001703:Biophysics / Q000379:methods; D002135:Calcium-Binding Proteins / Q000737:chemistry*; D002138:Calibration; D002846:Chromatography, Affinity; D003902:Detergents / Q000032:analysis* / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D005960:Glucosides / Q000737:chemistry; D007091:Image Processing, Computer-Assisted; D008565:Membrane Proteins / Q000737:chemistry; D008853:Microscopy / Q000295:instrumentation* / Q000379:methods; D018715:Microscopy, Video; D009004:Monosaccharide Transport Proteins / Q000737:chemistry*; D033902:Periplasmic Binding Proteins / Q000737:chemistry*; D011522:Protons; D013499:Surface Properties; D013501:Surface-Active Agents / Q000737:chemistry
D002135:Calcium-Binding Proteins; D003902:Detergents; D005960:Glucosides; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D033902:Periplasmic Binding Proteins; D011522:Protons; D013501:Surface-Active Agents; C023550:galactose-binding protein; C040358:dodecyl maltoside