The gene I'm working with in Drosophila melanogaster has orthologs in human as well as mouse. There are many commercial antibodies for these orthologs from different companies like Abcam, Sigma-Aldrich. I found an antibody recognizes a region with 90% similarity between Drosophila and human polypeptides. This antibody is polyclonal antibody produced in rabbit. The second antibody I found only recognize another region with 80% similarity between two orthologs. Since I'm not very familiar with antibody area, my question is what should be a good range of sequence similarity for antibody cross-reactivity? will that range depend on from which animals (mouse, rabbit...) that antibody is made?
Thank you in advance
Hi Nhan,
Do you want to use the same Ab for Dmel, Human, and Mouse tissues? If yes, and if the Ab is raised in rabbit, with 90% homology to recognition epitope across these species, it should work fine. You can choose the secondary Ab for eg. raised in chicken, anti rabbit, and avoid Sec. Ab raised in mouse.
Hope it helps. All the best.
Just be sure that the exact sequence of the peptide used to generate the antibody has that homology. Some suppliers will only state the approximate region where the immunizing peptide is picked.
But a 90% homology should be fine with a polyclonal
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