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removal of SDS from protein


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#1 anasyida69

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Posted 08 December 2009 - 12:59 AM

Does anyone know how to remove SDS from protein?

I run SDS-PAGE and cut my protein. I need the protein without the presence of SDS. Other than using electro eluter or dialysis, any other ideas?

Thanks

#2 mdfenko

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Posted 08 December 2009 - 07:21 AM

in the gel? or in solution?

either way, triton x-100 can displace sds and possibly renature proteins.

it is used in dialysis of solution against or by soaking the gel in a solution of (0.1%?) triton in an appropriate buffer.

Edited by mdfenko, 08 December 2009 - 07:22 AM.

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genius does what it must
i do what i get paid to do

#3 anasyida69

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Posted 08 December 2009 - 06:23 PM

the sample is in the gel. actually, i want to inject the protein into rabbit for antibody production. can triton-x completely remove sds? i have read the use of cold acetone, but don't know which one is good.

nway, thanks for the help.

#4 mdfenko

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Posted 09 December 2009 - 08:33 AM

we just homogenized the gel slice and injected (with adjuvant, etc). we didn't bother to remove sds or coomassie. antibodies were excellent.

triton displaces the sds and allows the protein to refold so that you may regain normal biological activity.

i haven't tried acetone for sds removal but used to use cold acetone to precipitate proteins.
talent does what it can
genius does what it must
i do what i get paid to do

#5 anasyida69

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Posted 09 December 2009 - 05:15 PM

yes..i agree with you. previously i did the same way too..sliced the gel and injected. but somehow i just afraid of the sds.

many thanks for your help..i will try it then :P




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