Does anyone have experiences to separate 2 to 5 KDa protein in SDS-PAGE procedure? Thank you.
Hi Vincent,
you can try by using a Tricine-SDS gel. You can find more information here:
http://www.ncbi.nlm.nih.gov/pubmed/17406207
If you can't buy precast gels (the best choice) here you can find a receipt:
http://www.oardc.ohio-state.edu/stockingerlab/Protocols/Tricine%20Gels.pdf
Yes you can try with Tricine SDS-PAGE to separate the proteins of low molecular weight as suggested by Alberto.
You can follow this paper
http://www.nature.com/nprot/journal/v1/n1/pdf/nprot.2006.4.pdf
Best wishes
Hi Vincent,
Tricine-PAGE gives best separation for smaller size proteins, due to its difference in pore size compared to SDS PAGE.
Check the nature paper suggested by Ashok.
Hi Vincent,
I don't know if you are gonna continue to transfer those small proteins to a membrane for WB. But in my lab, the general consensus is that, with small proteins, PVDF membranes do a better job than nitrocellulose. Specially small pore membranes, for us .2 uM pore membranes seem to work the best for ~10 kDa and highly positive charged proteins...
Thanks, all of you, I ought to evaluate those protocols you suggested. Hope everything goes well.
Dear Vicent:
Other opion is gradient SDS-page (for example between 25-15% acrylamide) or 2D SDS-PAGE.
Good luck,
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