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Detergent concentration: Difference between revisions
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This article is about how to measure detergent concentration after concentrating a membrane protein sample. | |||
1. The simplest way to control the detergent concentration is to use a | 1. The simplest way to control the detergent concentration is to use a | ||
higher cut-off concentrator if | higher cut-off concentrator if your protein plus detergent micelle is large | ||
enough. Michael Matho: “a 50kDa cutoff withheld a lot of detergent during | enough. Michael Matho: “a 50kDa cutoff withheld a lot of detergent during | ||
concentration process and consequently your final concentration might | concentration process and consequently your final concentration might | ||
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the “maximal cutoff you can use w/o loosing your membrane protein in the | the “maximal cutoff you can use w/o loosing your membrane protein in the | ||
flow through”. | flow through”. | ||
2. Patrick Loll, Edward A. Berry and John K. Lee suggested TLC, which | 2. Patrick Loll, Edward A. Berry and John K. Lee suggested TLC, which | ||
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increase sensitivity, speedvac a volume too large to spot on the plate, | increase sensitivity, speedvac a volume too large to spot on the plate, | ||
dissolve the residue in MeOH.” | dissolve the residue in MeOH.” | ||
A strategy for identification and quantification of detergents frequently | A strategy for identification and quantification of detergents frequently | ||
used in the purification of membrane proteins. Laura R. Eriks, June A. | used in the purification of membrane proteins. Laura R. Eriks, June A. | ||
Mayor, and Ronald S. Kaplan. Analytical Biochemistry 323 (2003) 234–241 | Mayor, and Ronald S. Kaplan. Analytical Biochemistry 323 (2003) 234–241 | ||
3. For sugar-based detergents (maltosides and glucosides), one can use | 3. For sugar-based detergents (maltosides and glucosides), one can use | ||
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glycosidic and bile salt-based detergents: applications in membrane | glycosidic and bile salt-based detergents: applications in membrane | ||
protein research. Urbani A, Warne T. | protein research. Urbani A, Warne T. | ||
4. Christopher Law: Use surface tension properties and look at the drop | 4. Christopher Law: Use surface tension properties and look at the drop | ||
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infrared spectroscopy). “very accurate, fast (10min) and requires as low | infrared spectroscopy). “very accurate, fast (10min) and requires as low | ||
as 10uL of protein sample.” | as 10uL of protein sample.” | ||
PVeesler, D. et al. Production and biophysical characterization of the | PVeesler, D. et al. Production and biophysical characterization of the | ||
CorA transporter from M. mazei. Analytical Biochem. (2009). 388 :115-121. | CorA transporter from M. mazei. Analytical Biochem. (2009). 388 :115-121. | ||
8. | 8. Philipp Ellinger recommended a nice new method that uses a fluorescence dye, Hoechst 33342, to detect micelle formation. The method is fast and easy, can be adapted to high throughput, and is not limited to sugar-based detergents. | ||
Jumpertz T, et al. High-throughput evaluation of the critical micelle concentration of detergents. Anal Biochem. 2011, 408(1):64-70. | |||