Protein extraction from plant leaves, bioactivity - (Oct/30/2009 )
My lab is trying to extract proteins from plantleaves and after extraction stimulate MCF7 cells with these extracted proteins.
We use a rehydrating buffer for the proteins containing 8M Urea. Does anyone knows an other rehydrating buffer for plant extracts which keeps the proteins in their bioactive state? Urea is not the best choice...
Hope to hear from you, thanks in advance.
I've extracted proteins from dried leaves, dried roots and dried seeds and never bothered with rehydration. I simply used aqueous solutions like phosphate or Tris buffers, salt solutions, buffered salt solutions (eg. PBS, TBS) etc. I assumed that if protein is insoluble then it's denatured and not bioactive anymore and useless to me. I had plenty of material to work with and never checked for efficiency, though...
Friend of mine was rehydrating leaves with glycerol solutions (around 1-10% as good as I remember) but it was not for protein extraction.
As for use of urea or guanidine hydrochloride - you could use some refolding method or protocol after that.