Need technique to know which protein isoform is expressed in Dro. muscle?

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S.A.D.
S.A.D.'s picture
Need technique to know which protein isoform is expressed in Dro. muscle?

I'm working with the Drosophila larvae as my model system.

My gene of interest makes five predicted polypeptide isoforms and I am trying to determine which of these isoforms is being expressed in the larval muscle cells. All these isoforms differ from each other in only the C-terminus, otherwise they have identical sequences. Their MW are in range of 115-130 kDa.

Does anyone have a good idea of what technique I can use to identify which of these five isoforms is being expressed in the muscle?

I tried Western Blotting, but the Ab we have recognises the N-termius, and it doesnt discriminate one isoform from another. I just see one single band. Someone else had mentioned PCR but I dont know how exactly to go about it.

Anyone have a good strategy and/or protocol?

Thanks

cfish
cfish's picture
Here is a link to a reference

Here is a link to a reference that may help on this subject. Methods are discussed and many references.

Dystrophin is required for
appropriate retrograde control of
neurotransmitter release at the
Drosophila neuromuscular junction

https://openaccess.leidenuniv.nl/dspace/bitstream/1887/12577/7/03.pdf