Anti-Estrogen Related Receptor beta antibody (ab19331)

Thank you for your enquiry regarding ab19331.

I can confirm that this antibody has been characterized for IHC-Fr (on frozen tissue), ICC/IF (on cultured cells or cytospin preparations) and in recognizes Estrogen Related Receptor beta in mouse. This receptor is expressed in ovary and the customer may wish to check the following website for further information on the expression profile:

http://www.ncbi.nlm.nih.gov/UniGene/ESTProfileViewer.cgi?uglist=Mm.235550

I would advise your customer to do some literature search (PubMed) to find out what sort of studies have been carried out specifically on murine ovary to monitor developmental expression of Estrogen Related Receptor beta.

If you need any further assistance in the future, please do not hesitate to contact me.

Thank you for your enquiry regarding our Estrogen Related Receptor beta antibody (ab19331) I can confirm the western blotting procedure used for testing this antibody was as follows: 1) Block membrane with 5 ml of blocking buffer for 1 hour at room temp., gently agitating solution. 2) Incubate primary antibody diluted 1:250 in blocking buffer for 1 hour at room temp. gently agitating solution. Peptide Pre-incubations: Incubate primary antibody in 100 ul of blocking buffer for 15 min. at room temp with peptide prior to incubation with filter. 3) Wash filter 3X with TBST buffer, 8 min. at room temp. each, gently agitating solution. 4) Incubate secondary antibody diluted 1:30,000 in blocking buffer for 1 hour at room temp. gently agitating solution. 5) Wash filter 3X with TBST buffer, 8 min. at room temp. each, gently agitating solution. 6) Visualize by ECL detection and expose film. Good positive controls for this antibody include: Rat kidney, testes and skeletal muscle. Unfortuantely, I am sorry we do not have a blocking peptide available for this particular antibody. I hope this information is helpful. Should you have any further questions, please do not hestiate to contact us again.

I'm very sorry to hear you are experiencing problems with ab19331, ab12986 and ab12987. I have looked at the shipping times of your order and would like to reassure you that there were no delays in the shipping. It is not uncommon that antibodies arrive at room temperature, especially when they travel some long distances and we have done stability studies in our laboratories and found that activity will not be impaired by such conditions. A high background problem is often due to the blocking agent or the secondary antibody; in our laboratory we block with 5% BSA (1hr) and find this buffer gives us the best results, while milk can give background problems so I think switching to BSA will help you a lot. Have you also tested if the secondary antibody works well with other primaries and if alone(no primary)it gives you some staining? For ab19331 the antibody has been tested in rat kidney samples and therefore I would like to recommend running this positive control as well as doing a nuclear extraction from your samples to concentrate the protein as you may have low levels of protein (you do not mention what cells you are studying), as well as incubating the antibody overnight in TBST. The antibodies ab12986 and ab12987 have not been tested in WB and therefore it is very difficult to know if those antibodies should work in this application and I would recommend trying the same protocol advice given above for ab19331. These antibodies have only been tested in human tissue so a human positive control is essential (e.g testis tissue). I think the above suggestions should help you a lot, if you still experience problems please do not hesitate to contact us. Ab19331 is guaranteed to work in WB in rat samples and if you still experience problems with this antibody in this application and species with the above changes I can offer you a replacement or refund,