about antibody Extinction Coefficient - (Oct/29/2010 )
I have a question about Extinction Coefficient.
I use the tool on http://www.expasy.ch/ and the software calculate the Extinction Coefficient and absorbance (optical density) ,using the following formula: Absorb(Prot) = E(Prot) / Molecular_weight. The Molecular weight in the formula is deduced from the amimo acid sequence I input. However, my sample is a kind of monoantibody which has two glycosyl-chains, the actual MW is bigger than the deduced MW. The software may forget this modification! Now I am confused and I don't know which MW should be used as Molecular_weight in the formula, the deduced value or the actual one? And I am also doubt if the glycosylation has effect on Extinction Coefficient.
Read some of the posting under "Antibody Concentration" at the top of the list...it may be of help to you.