Hi,
I am planning to test histone binding proteins (whole protein and different domains) using a Histone peptide array to determine the location and their modification(s) required for binding. I could not find a protocol to strip the array for re-use yet. As far as I found out they were mainly tested with antibodies where the binding strength is much higher compared to histone binding proteins. Has anybody experience with such an array and multiple uses?
Thanks in advance!
I'd try it, it should work like reprobing membranes for Western blotting. If you have a stripping protocol for antibodies, it also should work for proteins which do not bind so strong.
Maybe start washing with 50..100mM HOAc/0.9% NaCl or 50..100mM Glycin/HCl/0.9% NaCl pH 2.5, or 10..100mM NaOH/0.9% NaCl, then check if there is any signal left. Then validate the functionality of the array with the original sample.
If that works, find out how many times you may reprobe the array.
Thanks for the suggestions. I will test the different washing solutions.
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