protein inclusion bodies.. purpose of multiple water washes after lysate centrif - (Jun/22/2008 )

hi all.. for recombinant protein expressed in e.coli.. as protein inclusion bodies.. wat is the purpose of multiple water washes after lysate centrifugation?

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I never do that step where did you find this protocol ?

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most protocols have it.

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Never done it but I would expect that it's made to wash residual cytoplasmic proteins and maybe prepare the inclusion bodies to be lysed since the water would probably create an osmotic schok by changing abruptly the salt concentration

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The washing probably won't have that kind of effect on the IBs. They are hardy bu**ers, and usually require denaturation and sometimes reduction to dissociate them.
I would guess the main point is to wash off other proteins.

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HI

U probably mean washes with PBSx1 and not with water, it does not make any sense.

I also add 0.5- 1% of TRITON-x100, it helps retain and get rid of soluble undesired proteins from

the I.B.

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