Generally we use monoclonal and polyclonal antibody during western blot based on which works best for for us, its price and availability. But let's assume a scenario where a reference protein is being tagged with polyclonal antibody and target protein is by a monoclonal antibody, will there be difference in number of antibody bound to both the protein as reference gene will have total epitope = protein number x epitope per protein, while in reference gene single epitope will be present it should be equal to the number of protein.
Hi Sonti,
Indeed, polyclonal antibody preparations can recognize multiple epitopes whereas
monoclonal antibodies bind to a single epitope.
The question is whether you are interested in qualitative or in quantitative read outs. For
the
latter you need a titration of a reference and the target protein with known concentration (or
arbitrary units) to establish a signal to concentration relationship.
Keep in mind that the affinity of the corresponding antibody preparation is the most decisive
factor for the sensitivity/differences in sensitivity of your assay
kind regards
Urs
- Badges
- Science topic
- Similar topics
- Biological Science
- Anatomy
- Tissue