total protein extraction???

Supan ssupan at po-box.mcgill.ca
Thu Jul 20 13:32:31 EST 2000


Hi! Everybody:

I am new to this newsgroup. I am interested to know about buffers used to isolate total proteins from leaves of plant (i work with soybean and maize plants)? I am interested to measure the activity of following enzymes using the total protein (crude), i.e. Phenyl ammonia lyas, catalase, chitinase, glucanase, peroxidase. Anybody know a single extraction buffer to isolate total protein and check the enzyme activity. Furthermore, could someone please let me know whether it is feasable or not if i collect the leaves and gound them in liquid nitrogen and then free the powder at -80 C and then take the powder and dissolved in different buffer for differ enzymes? Do i need to freez dry the powder after crushing the leaves in liquid nitrogen or its ok if i just keep it at -80? If i use 80% ammonium sulfate, will i lose protein too? Any suggestion regarding total protein isolation will be greatly appreciated. Kindly send your suggustions to me at mwkhan at hotmail.com.


Thanks,
Wajahat Khan
mwkhan at hotmail.com
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