发信人: smileface (笑脸), 信区: Biology
标 题: Re: question on E coli protein expression
发信站: The unknown SPACE (Fri Jul 18 16:59:18 2003), 站内信件



for proteins like to stick in inclusion bodies, I strongly recomment
my protocol (in fact, from a literature I can't find the place leh)

http://www.sit.wisc.edu/~sguang/protocols/index.html

The last one is "using Sarkosyl to purify GST fused proteins"

u can try it.




【 在 sleepworm (hz) 的大作中提到: 】
: Every time I was trying to purify proteins, whose pI is either very low (<5.0)
: or very high (>10.5), they ended up in inclusion bodies. I am using pET28. So
: the proteins are His tagged and the strain I am using is BL21(DE3). The pH of
: my buffer is around 8.5. I have no problem with proteins with pI from 6-9.5.


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