I am working on Drosophila melanogaster and would like to know what would be a simple/easy protein that is known to associate with polysomes. The point of my experiment is to do polysome analysis, collect fractions and investigate which pool of fractions my protein of interest is associated with using western blotting. I am looking for a positive control - i.e. a protein which is known to associate only with 80S ribosomes and polyribosomes. I don't have antibodies for any large-subunit ribosomal proteins and so would like to look at other available alternatives.
Thanks Chi Zhang. However, I was looking for something which is not an RPL protein.
The standard approach would be to do your fractionation and collect fractions that correspond to your A260 tracings, do a TCA precipitation, and then perform westerns on the fractions. mRNAs that carry, one or two, vs. multiple ribosomes might not really have a differing protein content, just different numbers of ribosomes. You could try the Non-phospho-4E-BP1 (Thr46) antibody to indicate very inefficiently translating fractions, but this would likely be a marker of non-translating mRNAs. See Figure S2 in supplemental downloads: http://www.sciencedirect.com/science/article/pii/S0896627312001869
I think even eIF4E tends to fall off during polyribosome fractionation.
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