nobody know this! - (Sep/26/2007 )

hello every one,
i have separated IgG molecule and i runed on SDS-PAGE i have got 2 bands one at 25 kda that is light chain and 50kda ie heavy chain. now i need to separate light chain from each other.can i know which is the best technique for separation of thes light chain which have same molecular weight

thanks,
alie

-alie-

You need to do an affinity purification using antigen or peptide derived from this antigen first.

-genehunter-1-

QUOTE (alie @ Sep 26 2007, 11:29 PM)

Why do you want to separate the two light chains? I presume they are from antibodies with different specificities. What are you hoping to be able to do once you have done this?

-swanny-

QUOTE (genehunter-1 @ Sep 26 2007, 05:48 AM)

You need to do an affinity purification using antigen or peptide derived from this antigen first.

thanks for u r reply gene hunter and swanny,
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. now my problem is to separate the 2light chains ,which have same molecular weight.is their any technique to separate this?

alie.

-alie-

QUOTE (alie @ Sep 27 2007, 06:09 AM)

QUOTE (genehunter-1 @ Sep 26 2007, 05:48 AM)

You need to do an affinity purification using antigen or peptide derived from this antigen first.

you cannot separate polypeptides with identical properties, neiter by 1D SDS-PAGE nor 2D GE;

what do you like to demonstrate?

-The Bearer-

hi thnk u bearer,
i need to separate 2 light chains thats it.
alie

QUOTE (The Bearer @ Sep 27 2007, 09:44 AM)

QUOTE (alie @ Sep 27 2007, 06:09 AM)

QUOTE (genehunter-1 @ Sep 26 2007, 05:48 AM)

You need to do an affinity purification using antigen or peptide derived from this antigen first.

you cannot separate polypeptides with identical properties, neiter by 1D SDS-PAGE nor 2D GE;

what do you like to demonstrate?

-alie-

the light chains are identical. they are the same. you cannot separate them.

-mdfenko-