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Secretory protein?


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#1 johnny.jeng03

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Posted 28 July 2009 - 07:56 AM

Dear all:

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.

#2 genehunter

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Posted 28 July 2009 - 08:27 AM

Have you tried WB with concentrated and dialysed samples from the culture medium?

#3 johnny.jeng03

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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.




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