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Expression and Purification: Tips and Tricks: Difference between revisions
Expression and Purification: Tips and Tricks (edit)
Revision as of 16:12, 3 March 2008
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* 8. Dry pellet on a heat block at 95°C for 5-10 minutes to drive off acetone. | * 8. Dry pellet on a heat block at 95°C for 5-10 minutes to drive off acetone. | ||
* 9. Add loading buffer to sample and boil before loading on gel. | * 9. Add loading buffer to sample and boil before loading on gel. | ||
== Protein precipitates? == | |||
To increase solubility, follow Golovanov et al. 2004. In short, | |||
* buy L-Arg and L-Glu | |||
* put L-Arginine powder into water, start dissolving it, then add L-Glu powder, stir; add water to adjust volume as required | |||
* max. solubility is 1 M. The pH of a 1M stock solution will be around 6.4 or close to neutral | |||
* add stock solution (50mM) to your protein | |||