Posted 28 July 2009 - 07:56 AM
Recently, I have been investigating a novel protein in the fly Drosophila.
Based on following observations, I speculated it can be secreted by epithelium cells.
1. My protein is predicted to contain a signal peptide at the N-terminal.
2. Using immunostaining, this protein can colocalized with Rab7 (a late endosome marker) within cells that could be endocytosing it.
However, due to a lack of antibodies for secretory vesicles, I just wonder if I can use GFP-tagged Golgi marker instead.
If this protein indeed colocalized with Golgi, can I say that it is been secreting?
Moreover, what other experiments should I do in the future, in order to convince other that this protein is indeed been secreted?
Thanks in advance for all your advice.
Posted 28 July 2009 - 08:27 AM
Posted 28 July 2009 - 07:15 PM
Have you tried WB with concentrated and dialysed samples from the culture medium?
Yeah, I had a plamid containing C-termianl V5 tagged version of that protein.
I will try what you say in the future.