I recently used the ALP Assay Kit (Colorimetric) for the first time. I used it for culture media (and not cell lysates). However, I have 2 main issues:
1. The background value refers to my culture media that has phenol red (DMEM low glucose supplemenetd with 15% FBS), and its absorbance value is high, so that when I substracted it from the absorbance values of my samples, the values were negative.
2. The second main concern is that even with the naked eye, the color of the media of my samples were already different, which means that the background does not really represent that specific background of every sample.
I would really appreciate any suggestions and recommendations. I know that using cell lysates is better, but I just want to know if there is any way to measure ALP activity without the need to destroy cells.
Thank you in advance.
Hello Farah,
I am also using ALP kit for quantification,
Each sample should be it's own blank, prior to starting the assay place the same test voulme of media to the additives (pNPP without ALP enzyme) to empty wells ext to each sample, incubated for the same time under same conditions, read for absorbance and subtract to get your meausrments.
Dear Ahmed,
Thank you very much for your reply. I will hopefully run the test again and I will follow your instructions.
Best,
Farah
- Badges
- Science topic
- Similar topics
- Social Psychology
- Attitude