We identify an IgM antibody from human, which specifically binds to the outer membrane of breast cancer cell line MDA-MB-231 and is good for CDC in this cell line.Is anyone thinks it is interesting? Could you help me to figure out the ways to find its target? From westernblot it appears that it recogizes a 170KD protein.
Dear Huiwu,
A very potent way is by peptide mapping/mass spectrometry. For this you need to enrich and partially purify the target, e.g. by immuoprecipitation with the IgM, or bind your IgM to a column and try to purify the target, etc. Then confirm by western blotting that it is your target. Finally, identify the protein by peptide mapping/mass spectrometry.
See e.g. Rauschert, et al. (2008) A new tumor-specific variant of GRP78 as target for antibody-based therapy. Lab Invest 88: 375–386.
Good luck,
Andreas
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