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Thanks for the quick response. I am going to send one of my blot. Please tell me what you think. Also Why can't this antibody detect active form of MMP-7? For the information, we are using HT-29 human colon cancer cell line and the cells were transfected Syndecan-2 (heparan sulfate proteoglycan) and the Syn-2 induced MMP-7 mRNA level and the protein (if the band we have is right). Thanks for your help. |
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ANSWER: |
Thank you for sending me the image of your blot, I understand your question. As I previously mentioned, the predicted MW of the protein (precursor, prior to any post-translational modifications) is 29.6kDa. I have tried to find further information regarding the exact MW of the protein and would like to direct you to the Swiss prot data for human MMP7 http://us.expasy.org/cgi-bin/niceprot.pl?P09237 and to a SDS gel on this database showing the band slightly higher than 30kDa (SDS-PAGE of nucleolar proteins from Human HeLa cells) http://us.expasy.org/cgi-bin/ch2d-compute-map?NUCLEOLI_HELA_1D_HUMAN,P09237 I do not know why your gel does not show a lower band for the active form of MMP7, it is possible that your samples do not contain this form of the protein or low amounts (below WB detection levels). I would recommend running a positive control of conditioned, serum-free medium from (TPA-treated) human SW620 cells or Placenta, bladder, breast and ovarian carcinomas to confirm that the band seen is indeed MMP7. I hope this information helps, please let me know if you need further assistance,
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