Hi all,

I am trying to measure the beta-hexosaminidase release in HEK-293 cells expressing recognition receptors after stimulating them

I have tried p-NAG (4-Nitrophenyl N-acetyl-β- D -glucosaminide) assay and fluorescence assay and for both, I had lysed the cells with Triton 0.1 to know the highest B-hexo release. In both assays, I did not see any significant difference between control and stimulated cells.

I follow the 4-Methylumbelliferyl N-acetyl-b-D-glucosaminide protocol with 2mM concentration for substrate in 0.05 M citrate.

Could you please help me troubleshoot it

Thanks