Hi Everybody, I was wondering if anybody had a similar case to mine. The protein I'm working on is a tough one. Its around 75 KDa in size, and it purifies well, but soon after the full length protein starts to autolyse. Consequently, I don't get any hits after setting up the screens.
To get around this I digest the protein with trypsin and end up with a stable fragment of around 25KDa. When I screen this I get many hits, but none of them look like something that can be made into single crystals. All the hits give me large lobe-like clusters and they look like cauliflower heads but they are optically active. I currently have the protein in 300mM NaCl buffer. Would dropping or increasing the salt or even using a different salt make any difference? Any ideas? Cheers, Chris
