coating Serum - (Sep/04/2008 )

Hi,
I know it is a weird question, but is it possible to coat human serum to detect a protein in it? We have a couple of antibodies (monoclonal and polyclonal) but a sandwich just doesn't seem to work....

Greets

you will have competition between your protein of interest (probably in low concentration) and other major proteins in the serum for attaching to the wells of your microtiter plate. Examine how hCG, TSH, LH, TnI, and other protein markers are detected and follow their lead.

coat the wells with your ab!

good luck

hi,

if capture ELISA is not working for ur protein of interest. take the antibody which ever can recognize the target protein in the serum and pull down with protein A beads (ofcourse u will pull down lot of serum antibodies along with ur target Ab). perform western blot with same antibody used to pulldown ur sample and observe the results.

gud luk