Hello everyone,
I am working to isolate/immunoprecipitate homologs of a protein (RdR) from an invertebrate. There are no commercially available antibody for any of the homologs for my organism. However, there exist a polyclonal antibody against another homolog of the protein in C. elegans. Since the two proteins are related and predicted to have similar function (this have not been experimentally verified), is it possible that I can obtain the said polyclonal antibody and use it to immunoprecipitate my protein of interest? and what things do I need to consider if I should/shouldn't go this route?
Is there anything you guys suggest I do?
Also, I'd like to point out that the predicted targets (epitops) by the company are not a 100% match to the homologs I am working with as seen below from multiple sequence alignment.
If the question isn't clear enough I can explain more. Thanks in advance
Jacob,
If you're lucky, it works. It largely depends on the grade of sequence homology. Do you have an opportunity to compare the protein sequence of your target with that from the C. elegans protein?
Yes, on of the six homolog appear to share some sequences with the one in c.elegans but not a 100%. probably < 50% in the epitop region. see updated sequences in question above @Wolfgang Schechinger
Jacob, that's very hard to read. Multalin is great tool for generating colour coded alignment: http://multalin.toulouse.inra.fr/multalin/
But with 50% homology on AA basis, I think chances for a specific signal are low. If you can't find a suitable antibody, consider having your own made against a selected synthetic peptide sequence. Esp. when you anticipate that you'll need larger amounts. A polyclonal is fine for that.
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