Hi Sandra, >From your post, it looks to me like you are assuming the recommended dilution should work. I apologize in advance if I have misinterpreted your post in this regard.
When starting with a new antibody (including a new vendor for same Ab and/or a new clone), do a range of dilutions that includes the recommended one. So in this case, I would try 1:50, 1:100 and 1:200 to start. Use the recommended AR first and then see how each of the 3 dilutions look. Just as an example, you may find 1:50 has a weak signal and higher dilutions are negative. If so, set up 3 more dilutions with the same AR (eg 1:10, 1:25 and 1:50). Once you get close to where you think the intensity should be you can try modifying the AR protocol to improve the sensitivity. Good luck, Greg -- *Greg Dobbin* 1205 Pleasant Grove Rd RR#2 York, PE C0A 1P0 *Everything in moderation...even moderation itself**!* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
