IgG separation
nishanth nataraj
Thursday, 27 September 2007 13:11 UTC
hello every one
i have done affinity purification and i have got pure IgG moleucle.i runned on the SDS-PAGE i have got bands according to size thats is 25kda-light chain and 50kda heavy chain. now my problem is to separate the 2light chains ,which have same molecular weight.is their any technique to separate this?
nishanth.
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Replies
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Hi Nishanth,
the problem you have is that in any one whole IgG molecule the light chains are identical. You could separate the light chains from different IgG molecules in a purified preparation of polyclonal antibodies by using 2D-SDS PAGE. This may prove difficult if they have a similar isoelectric point.
It does also depend on why you need to separate them because other techniques such as HPLC may be useful too, particularly if you need to sequence the chains or use them for other down stream applications.
Hope that helps!
Mikayala -
hello King,
thank you very much for u r reply.
nishanth
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