How much Antibody did I get? - Protein G column (Mar/21/2006 )

Hello-


there is std fomula to estimate the pure Antibodies
if it is IgG then
mg/ml=OD at 280/1.5

first take the blank reading (the buffer in which ur Antibody is there) at 280 and take ur sample reading and deduct the sample reading form blank,then apply the formulae u will get the concetration of ur antibody

best of luck

I tried purifying antbodies using a Protein G column from hybridoma supernatants for the first time yesterday.

After elution in acidic glycine I got two 1ml fractions that absorb at
~A280=0.100

Any idea how much antibody that is or how to convert it into a protein concentration roughly? I used a pretty small column thinking I might screw up my first time and only 80mls of supernatant so I'm not expecting much. I never use our spec and have really no idea what 0.100 means. Any ideas?
I will do a protein assay in a couple days after dialysis but wouldn't mind knowing about how much I have sooner if possible so that I can purify more if necessary.


Thanks,

Mountainman