Dear Colleagues.

Hi. We did immunostaining in some canine samples containing fat to detect adipokines. We pre-incubated the samples with 5% BSA to reduce the background staining, however our background was too intense. We also used EDTA 1 mM (PH 8.0) as Ag retrieval solution.

1- Any idea to get better results if I swap to another blocking solution for non-specific background staining? If so, please let me know what solution?
2- Will the results improve if I switch to citrat buffer instead of EDTA as Ag retreival solution?

Any comments will be appreciated.

Thanks for your consideration.

Siamak from Australia

Siamak,

Results vary with the antigen retrieval method. You'll have to try a variety of methods and anitbody concentrations to find what works for you. For some ideas read this site.

http://mousepheno.ucsd.edu/immunohistochemical_prototype.shtml

Rb

Good Luck let us know what you find out