We have tried using normal colon as controls but lymphocytes are staining as well. Getting a weak cytoplasmic staining as well. Tried with and without HIER and various dilutions.
you need to specify what kind of assay you plan to use. Very often the abs that are good for IHC do not work for with fluorescent microscopy or flow cytometry etc. Lots of Abs do not work in human tissue simply because there is no expression of the protein or the protein is modified/mutated etc.
One of the best ERbeta Abs for human tissues are from Dako clone PPG5/10 -
http://www.agilent.com/en/products/immunohistochemistry/antibodies-controls/primary-antibodies/estrogen-receptor-b1
the Dako Abs gave good results with human tissues - look at the published data
http://clincancerres.aacrjournals.org/content/23/3/766.long
Abcam Abs ERbeta clone EPR3778 are also good -
http://www.abcam.com/estrogen-receptor-beta-antibody-epr3778-ab133467.html
Abcam sells lots of different abs for ERbeta. 14C8 are not bad, we used them and they worked ok; read the customer reviews before buying and check the PubMed for the papers that used the ERbeta abs.
One of the tissue positive control are MCF-7 and/or T47D mammary cancer cells, but lots of people also use endothelial cells
I used Thermo PA1-311 for Western blot. Works fine and discriminates ERbeta from ER alpha. You should know that lymphocytes express numerous isoforms of ERa and ERb (PMID: 19608332, tables 1 and 2). So what you see may not be an artifact. We checked specificity by transfecting HEK cells with various isoforms. I see a lot of ERbeta and ERalpha-delta4 in the cytoplasm of MT-2 cells - a model of Tregs.
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