I am getting a band when in both acetyl CoA treated and target protein. I think Acetyl CoA itself showing the acetylation. So, I cannot demonstrate my target protein have the potency of acetylation. I am confused as to confer my protein has the property of acetylation.

5X acetylation assay buffer：0.25M Tris-Hcl(PH:80), 0.5mM EDTA, 5M DTT, 50%Glycerol.

(0.5 μg) was incubated with (0.5 μg) in an acetylation buffer (50mM Tris/pH 8.0, 0.1mM EDTA, 1mM dithiotheritol,

10mM sodium butyrate, 2mM acetyl CoA, and 10% glycerol) at 37 °C for 0.5h, 1h.

I am getting

Please, can suggest to me any condition to overcome this query.

Thank you in advance.