Anti-DDX3 (phospho T322)抗体(ab61043)

Thank you very much for your answer. As my colleage is out of the office today, I am forwarding you already the answer from the laboratory in regards to the specificity of the ab61043. Please find the answer here below. I would like to reassure you again, that we do cover all our products with the guarantee to work as stated on the datasheet. If therefore the suggestions of my colleague will not help to resolve this issue, we will replace or refund this antibody for you. Can you please also confirm that the samples have been thoroughly denatured (with SDS and heat) and reduced for the loading?  Also, can you maybe send us an image of the results? Please find here the answer from the lab: " The immunogen peptide for this antibody has no significant homology with DEAD box domain. The name of the target protein (DEAD box protein) is causing confusion. (A)-This antibody was produced for phosphorylation site T323 of human DDX3X (We count phosphorylation site as T322, because the first M will be removed after the protein is synthesized).  (B)-Other name for this target is DEAD box protein 3.  (D)-By homologue search, the immunogen peptide only hit DDX3X.  (E)-The immunogen peptide is not located or has any significant homology in/to helicase ATP-binding domain, Q motif, and/or DEAD box, so this antibody will not react with any proteins containing these domains.  (F)-The reference for phosphorylation site: Biochem Biophys Res Commun. 2007 May 11;356(3):668-73. Epub 2007 Mar 15. " Please let me know how you are proceeding and whether you have any questions.  I wish you meanwhile a good weekend.

Thank you for taking time to complete our questionnaire. Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab61043 : 1) I would suggest to use the antibody in a tested application, ELISA or ICC/IF. This antibody may not be suitable for Western Blot, we do not have any experimental data for this application. Our products are not covered by our 6 months Abpromise guarantee for untested applications (or untested species). For more details about the Abpromise guarantee, please visit the following link : www.abcam.com/abpromise 2) I would recommend to use BSA as blocking agent and dilution solution for both primary and secondary antibodies. Milk is not recommended for studies of phospho-proteins because milk contains casein which is a phospho-protein; it may cause background because the phospho-specific antibody detects the casein in the milk. I hope these suggestions are helpful.

Thank you for contacting us. As far as we are aware, ab61043 has never been tested in western blot. All tested applications are specified on our datasheets, and these are updated as soon as any new information is brought to our attention. I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. How different is the oberved molecular weight from the expected one? I look forward to receiving your reply.