Hello,
I'm trying to inhibit bovine intestinal alkaline phosphatase in a supplied kit reagent. Since the components of the kit are proprietary, I don't know the actual concentration of alkaline phosphatase in the bottle so trial and error with various inhibitors is the only way I can test for inhibition.
Various papers in the literature have described using amino acids (tryptophan, leucine, phenylalanine) for inhibition of this enzyme. But I'm having trouble getting it to work. There are other enzymes in the reagent that I don't want to impact, so amino acids are ideal for this because I don't want to mess with the pH or other conditions requires by the other enzyme in the mix. I'm also having trouble getting high enough concentrations for inhibition when solubility of these amino acids in water is so low.
Has anyone worked with alkaline phosphatase inhibitors that can give me some advice? Alternative inhibitors to try? Concentrations to try? I just need some fresh ideas. Thanks!
Vanadate is probably your best bet.
Here is a source of references for alkaline phosphatase inhibitors.
https://brenda-enzymes.org/enzyme.php?ecno=3.1.3.1#Ki%20VALUE%20[mM]