Advice on primary antibody for detecting PDI1 in moths

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hesp
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Advice on primary antibody for detecting PDI1 in moths

Postby hesp » Wed Apr 01, 2015 1:44 pm

Hi there,
I recently conducted a study looking at protein disulphide isomerase (PDI1) expression/upregulation in moths. The primary antibody that I used in the membrane incubation after western blotting was Anti-PDI1 [38H8] antibody (ab4644) which is monoclonal, and most commonly used in various other organisms such as humans, rhesus monkeys, yeast etc. It was readily available in the lab I was working in and works for fruit flies, so I thought it may work for moths.
After transferring my polyacrylamide gels onto membranes via western blotting, the only banding I could see after staining the membranes with a substrate was from the kaleidoscope marker, despite using Saccharomyces cerevisiae (bakers yeast) as a control on my gels.
I just wondered if anyone had any interesting input as to why this might be (if the antibody is not specific enough, if the antibody may be too old, yeast not incubated for long enough), and if anyone is aware of any other antibodies which can be used to detect PDI1 in Lepidoptera or has had similar problems?
I'm finding it hard to find online journals related to this subject specifically.

Many thanks,

Hannah.

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