PharmPK Discussion - Antibodies to Carbonyl Reductase

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On 2 Oct 2006 at 12:25:47, "Gwyer Lovell" (Gwyer.Lovell.-a-.medivir.com) sent the message


Hi,

I need to cary out an experiment to assist identification of the
enzyme present in hepatic cytosol that reduces a compound I am
currently working on. My hypothesis is that one (or both) of the
human carbonyl reductases (CBR1 and CBR3) are implicated. I have
polyclonal antibodies to both enzymes (300ug of each).
I'm however at a loss as to what range of antibody concentrations to
use for co-incubation with 0.5mg/mL cytosolic protein and 2uM
substrate. A colleague suggested to nominate 4x cytosolic protein
(i.e. 2mg/mL - very expensive!) as my highest antibody concentration
and work down from there. Does anybody have any suggestions or know
of any relevant literature references?

Many Thanks, Gwyer

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