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BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER Nep90535 DESCRIPTION OF THE PROBLEM Non-specific band at 39kDa which is cell trype specific, and no strong band at correct MW SAMPLE human cell lysate DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED none used ANTIBODY STORAGE CONDITIONS aliquoted at -20 AMOUNT OF PROTEIN LOADED 50ug ELECTROPHORESIS/GEL CONDITIONS reducing, 10% gel TRANSFER AND BLOCKING CONDITIONS 5% blotto in all antibody incuabtions SECONDARY ANTIBODY santa cruz anti goat HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes ADDITIONAL NOTES Can you recommend a positive control for this antibody
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ANSWER: |
Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. The following publication makes reference to the use of rat ovary as a positive control for IHC and RT-PCR analysis of Activin Receptor Type IIA expression. Refaat BA, Bahathiq AO, Sockanathan S, Stewart RL, Wells M, Ledger WL. Production and localization of activins and activin type IIA and IIB receptors by the human endosalpinx. Reproduction. 2004 Aug;128(2):249-55. PMID: 15280564 http://www.reproduction-online.org/cgi/reprint/128/2/249 However, this antibody is yet to be tested on rat cells. Therefore from a literature search I have determined that the following publication used HHUA cells as ositive control cells in flow cytometric analysis. Tanaka T, Toujima S, Umesaki N. Activin A inhibits growth-inhibitory signals by TGF-beta1 in differentiated human endometrial adenocarcinoma cells. Oncol Rep. 2004 Apr;11(4):875-9. PMID: 15010888 http://147.52.72.117/or/2004/volume11/number3/667.pdf HHUA cells have been shown to express all four types of activin receptors in the following publication. Tanaka T, Toujima S, Otani T, Minami S, Yamoto M and Umesaki N: Expression and function of activin receptors in human endometrial adenocarcinoma cells. Int J Oncol 23: 657-663, 2003. I would appreciate it if you could tell me whether Activin Receptor Type IIA is subject to processing in the cell types that you have been using. What do you mean by a cell type specific non-specific band? Should you continue to have problems with this antibody please feel free to contact us again so that we can obtain further details off you such that we can make some recommendations as to how you might improve the results that you have been obtaining. |
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