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Smearing antibody and DTT - how to interpret the gel? - Aggregated antibody (Apr/16/2007 )

I am working with an anitbody that is highly glycosylated and tends to aggregate. When it is loaded on SDS-PAGE it is very "smeary" from the full IgG-band and all the way to the wells. I interpret this as if there are aggregates of many different sizes in the material. If I add DTT, the antibody is reduced to form only two bands, the heavy and the light chain, and there is no signs of any smear. My question now is if this means that there are "faulty" disulfide bridges formed that bild up the aggregates or what does this really mean?

Thanks in advance!
/B

-kenzo-

QUOTE (kenzo @ Apr 16 2007, 11:49 AM)
I am working with an anitbody that is highly glycosylated and tends to aggregate. When it is loaded on SDS-PAGE it is very "smeary" from the full IgG-band and all the way to the wells. I interpret this as if there are aggregates of many different sizes in the material. If I add DTT, the antibody is reduced to form only two bands, the heavy and the light chain, and there is no signs of any smear. My question now is if this means that there are "faulty" disulfide bridges formed that bild up the aggregates or what does this really mean?

Thanks in advance!
/B


glycosylation and disulfide bridges are different features; you may deglycosylate by incuabtion with N-/O-glycosidases

-The Bearer-