There is a protein has two isoforms, one of which has a signal peptide. I am wondering if it is a secretory protein. If it is, how can I detect it? What if it is secreted by extracellular vesicles pathway?
Thank you for your help!
Hi,
You can do the usual immunofluorescence on the sections/cell culture (depends on what you work with). Of course, if there are antibodies available which can distinguish those 2 isoforms. Alternatively, you can use techniques like in-situ sequencing, though they are much more laborious.
Best,
Michail
If your protein is secreted, the signal peptide will get cleaved off during import into the ER. if you have an antibody you can use for pull-down, you can use mass spec to look for the properly processed form, both in cell lysate and in conditioned medium.
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