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Plant protein extraction and purification - how to extract plant proteins and purify them? (May/29/2006 )

can someone tell me how to extract plant proteins n how to purify them?

thanks a million



there are lots of methods

i personally do a very simple one

weigh the plant leaves
use liquid nitrogen / freeze them in - 70 for a couple of hours

grind well using ice cold 1X PBS ( u can use 2mM PMSF or other inhibitors of proteinases )
transfer to eppendorff or centrifuge tubes
centrifuge at 10000 rpm for 10 min at 4 degree
collect the supernatant

this will be total protein

after this u can do acetone purification
add ice - cold acetone , 2- 3 times the volume of supernatant
keep at - 70 for 1- 130 hour
centrifuge again same conditions as previous centrifuge step
save pellet
wash the pellet with 70% cold ethanol
dry the pellet
dissolve in 0.1M Tris Hcl ( ph 8.3)

but then protein concnetration might get a bit lost

there are other protocols too

there are a number of kits available too