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Thanks for Support. Please find below replies to the questions you have. Hope they may assist. ab47409 - Anti-FOXO3A antibody- Order No: 946985 (1023048) - for Steve 3.1 Concentration: 1:500 Question 3.1 Have you tried different dilations such as 1/000? yes, but 1/1000 was not as good as 1/500. 3.2 Secondary antibody: rabbit Question 3.2 Could you please specify the host species of the secondary antibody used and what type of immunoglobulin it is specific to? rabbit IgG 3.3 Signal enhancing detection: Question 3.3 This antibody has been used for ICC/IF at 1 or 5 ug/ml using signal enhancing detection system (polymer-base The point is the antibody does not label the phoph-foxo3 that we have confirm exist in the cytoplasm with the antibody of p253-foxo3.
Asked on Dec 01 2011
Thank you for getting back to me and for providing some further information. It is much appreciated. This antibody is rabbit polyclonal so the host species of the compatible secondary antibody should be different from rabbit. Reading your response, it seems you are expecting to detect the phospho FOXO3A (at serine 253) with ab47409 . It is important to emphasize that as the datasheet indicates the immunogen used to raise ab47409 was a synthesized non-phosphopeptide derived from human FKHRL1 around the phosphorylation site of serine 253. Therefore this antibody only detects the non-phosphopeptide FOXO3A. For your information, we have some specific antibodies against different phosphorylated FOXO3A at different positions, but these product names and their specificity state that they only recognize the modified version of FOXO3A: ab47285:Anti-FOXO3A (phospho S253) antibody:https://www.abcam.com/foxo3a-phospho-s253-antibody-ab47285.html ab31109:Anti-FOXO3A (phospho S253) antibody: https://www.abcam.com/foxo3a-phospho-s253-antibody-ab31109.html ab26649:Anti-FOXO3A (phospho T32) antibody:https://www.abcam.com/foxo3a-phospho-t32-antibody-ab26649.html I hope this helps and if I can assist further, please do not hesitate to
Answered on Dec 01 2011