Anti-SATB2 antibody [SATBA4B10] (ab51502)

Hi,

I am testing your antibodies ab51502 and ab92446 for WB and IP and would like to know the concentration of the antibody.

Also, is there a recommended IP protocol or preference of the 2 antibody for IP or WB?

Thank you!

ANSWER:

Thank you for contacting us. The concentration of ab51502 is 0.1 mg/ml and should be used at a dilution of 1/100 for both IP and WB. Regarding ab92445, this is a tissue culture supernatant antibody so the concentration is not determined. Generally concentrations are only determined for purified antibodies as they are not relevant for other types that may contains proteins other than the protein of interest. This product should be used at a dilution of 1/10-1/100 for IP and 1/1000-1/10000 for WB. We do not test our antibodies side by side and cannot say which works "better" for a particular application but we do guarantee them both to work in IP and WB.



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Ab before worked well with other lot
this lot GR19837-3 gives no signal in IHC (PFA fixed) and WB
Ab: 1/500 - 1/200 (IHC)
1/1000 - 1/400 (WB)
same protocol, same samples as before

ANSWER:

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Dear Abcam, I have discovered that in fixed adult rat cerebral cortex sections, the mouse anti-satb2 antibody recognises both satb2 and satb1 proteins (IHC). Interestingly in mouse cerebral cortex sections, it binds to satb1 but not satb2 protein. Unlike the anti-satb1 antibody from another company, which seems to label weakly satb2 protein and strongly label satb1 protein in rat tissue, ab51502 strongly recognises both satb1 and satb2. I hope this is helpful to you,

ANSWER:

Thank you for bringing this to our attention. After reading your email I was able to perform an alignmet of the immunogen sequence used to create this product against mouse and rat SATB1 and SATB2 proteins. I found that there was62% homolgy between murine Satb1, Satb2 and the immunogen. This alignment isat the C-terminal region. This product is a rabbit polyclonal antibody, therefore it contains antibodies to a number of different epitopes along the immunogen sequence. While we never recommend an antibody that has less than an 85% homology to the target, it may be that in this case, because of the multiple epitopes, thatab51502 would react with Satb1 and react differently in the different species. However, I am concerned that ab51502 did identify SATB2 in mouse in IHC. I would like to help you fix this. Would you be willing to send me your protocol used for IHC, any troubleshooting that you may have done as well as any images or data that you obtained so that I might go over those? I may be able to offer tips or suggestions to help you or to replace the product if neccessary. I hope you have found this information helpful. Please contact me if you have any questions.

Thanks for your help.  I did finally get a signal after letting the exposure go for much longer, so it looks like it worked.  I would like to get a 2nd SATB2 antibody to use to confirm the IP I plan to do.  Can you recommend another SATB2 antibody that has been successful in Western analysis or IP?  

ANSWER:

Thank you for your email. I am very glad to hear that you obtained the desired results.

As for another SATB2 antibody that is suitable for IP/WB, we have also ab92446: http://www.abcam.com/SATB2-antibody-EPNCIR130A-ab92446.html It was tested in IP and WB and several other applications, as well as in human, mouse and rat samples.

The antibody you are currently using is also suitable for IP: ab51502

Other antibodies tested in WB and human samples are: ab69995, ab69998 and ab119118.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.