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 Protein purification from Inclusion Bodies [View Printable]
Pinal Pandya

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Hi there,
I am trying to solubilize my protein from Inclusion bodies. I used DH5 alpha strain for the expression. And i found this protocol online for protein refolding. It says i need to dialyse the protein using urea. But my problem is i dont know where to buy the dialysis bags from. I am stumped !!! Can anyone help me.
Thanks
Pinal
.........................

Posted Oct 12, 2007, 19:17 PM
R Bishop

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Pinal,

I find that Pierce has some nice dialysis products. I use the Snakeskin for large amounts of solution and the slide-a-lyzer cassettes for smaller volumes.

http://www.piercenet.com/products/browse.cfm?fldID=040101

Also trying using the Solution Search link on this page as there are many threads about inclusion bodies and refolding

Hope that helps

Rb
.........................

Posted Oct 12, 2007, 21:33 PM
Pinal Pandya

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Thank you soo much for your help RB , this definitely helps a lot and i am going to check out the solutions search link.

thanks again
.........................

Posted Oct 12, 2007, 21:41 PM
Pinal Pandya

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Thanks for helping me before RB. I have another question, I found this protocol online for solubilization and refolding. the method is as follows: add 100mM tris buffer ph 8, 50mM glycine ; sonicate; dissolve supernatant in the above buffer + 8M urea.

then add 5mM reduced glutathione and 0.5 mM oxidized glutathione. and then proceed for dialysis.

so i guess my question is why do we need ot add glutathione at all? reduced and oxidized? and if we use oxidized glutathione with ~6% ethanol , is it ok?

Please help me!!!
Pinal
.........................

Posted Oct 16, 2007, 17:43 PM
Pinal Pandya

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