We use a polyclonal antibody from R&D system produced in goat but we obtain several bands in our westerns (green bands). I show a picture, in that western blot, we applied 30 ug of total protein in a 10 % polyacrylamide gel and reduced conditions, the blocking step was with BSA. The red band is b-actin. The theoretical molecular weight is around 32 kDa, the receptor has potential glycosilation sites and it has been reported that it is glycosylated, but as you can see in the picture, the pattern of bands are in a very wide range and I suppose that the polyclonal antibody recognized other proteins. I have tried different antibody dilutions. The cleaner assays obtained was when I loaded 20 ug of protein, but the specific signal at 32 kDa was too low.
I would suggest you use a monoclonal antibody. It is not unusual for polyclonal antibodies to detect multiple bands or epitopes. Try the Boster product. It may work better for you. I found their site: http://www.bosterbio.com/anti-lox-1-antibody-pa1832.html. Good luck.
Thomas
I see no real problem if your polyclonal AB is detecting other proteins, as long you can be sure that your band of interest is that what you are looking for. If in doubt, have it sequenced by mass spec. If your polyclonal should be derived from a peptide antigen, you may expect the same result from a monoclonal and it might make sense to look at the other proteins it detects, too, as they might be isoforms.
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology
- Antibodies