Lysyl oxidase (LOX) activity kit problems in skin fibroblasts?

25 March 2021 1 1K Report

Hi, I am using the Lysyl Oxidase kit from AAT Bioquest (https://www.aatbio.com/products/amplite-fluorimetric-lysyl-oxidase-assay-kit-red-fluorescence).

My goal is to use this kit to measure LOX activity in conditioned media, however, when I do the blank with media alone (no contact with cells) it gives more signal that the conditioned media. I am working with human skin fibroblasts where the activity of the enzyme is high. I am using positive (cells should have LOX activity) and negative (cells should have lower LOX activity due to copper transport proteins KO) controls and they give the same intensity.

I tried DMEM media without red phenol in case it interfere with the measurement and nothing.

Any advice?

Thank you.

Kc Chico

Yes, my first thought would be the media as well, but if red phenol wasn't the issue, then I would still recommend exploring your options: https://www.aatbio.com/resources/cell-media-formulations/

If that doesn't help, I can ask our scientists what might be the issue.

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