We have in our lab all the basic equipments for protein purification, e.g french press, centrifuges, ultrafiltration dyalisis, ion exchange chromatography... The source organism should be preferrably non-pathogenic.
The plasma membrane P-type H+-ATPase from yeast and the F-type H+-ATPases from spinach chloroplast, beef heart mitochondria, and E.coli are all well-studied proteins that you could purify based on published methods. Remember that these are membrane proteins, so they are more difficult to work with than soluble proteins. You will need to extract them with detergent and perform all chromatography in the presence of detergent. The spinach enzyme is very convenient because you can use fresh spinach from the grocery store as the source.
Many thanks for the tips of the fresh spinach. I work extensively with the photosynthetic reaction centres, which are membrane proteins so I should be able to follow a protocol with all the necessary steps.
- Badges
- Science method