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Why does this antibody detect a doublet around 50 kDa? The other Zhangfei antibodies in your catalog detect bands of different molecular weights, and the predicted molecular weight of this protein is 38 kDa. Are there multiple isoforms of Zhangfei? What is this antibody detecting?
Asked on Oct 04 2011
Thank you for contacting us. I apologize for the delay in my reply. Our lab provided the following explanation for the WB data: This seems to be a challenging target; in Biochem. J. (2008) 416 (463–473) they state: “SMILE (SHP-interacting leucine zipper protein) was previously designated as ZF (Zhangfei). SMILE gene generates two isoforms [SMILE-L (long isoform of SMILE) and SMILE-S (short isoform of SMILE). ZF is only a partial clone of CREBZF, which encodes protein with a predicted molecular mass of 37.2 kDa. Since ZF is a member of the bZIP family, we named CREBZF as SMILE. (A) schematic of SMILE-L (CREBZF) and SMILE-S (Zhangfei/ZF). (B) Exogenous expression of human wild-type SMILE and ZF in HEK-293T cells using the pcDNA3 vector. WB with SMILE-specific antibody. (E) Expression of SMILE protein in cell lines (left-hand panel) and mouse tissues (right-hand panel). To summarize, there is a long isoform, 50–60 kDa (SMILE-L), and a short isoform, 40 kDa (SMILE-S = ZF), and our ab seems to recognize the SMILE-L form. I hope this helps to clarify the results. If you are not satisfied with this antibody, I would be happy to offer you a free of charge replacement, credit, or refund. Please let me know your original order number and how you would like to proceed and I would be happy to assist you further.
Answered on Oct 04 2011