Alkaline phosphatase (ALP) is an enzyme often measured as marker of activity of bone cells cultured in vitro. If the medium contains fetal bovine serum, could the ALP content of the serum affect the measurement of the cellular enzyme? Do you know the approximate concentration in FBS? Should it be subtracted from the cell ALP value (as background)?
Hello Gabriela Ciapetti
The effects of ALP activity intrinsic to FBS can affect experimental outcomes. Please refer to the article below for more information.
Preprint Alkaline phosphatase activity present in serum influences os...
I am providing a link below which carry out biochemical tests (alkaline phosphatase being one of the test) as part of their Quality Control Programme. You may get the information you require by contacting them.
https://www.bioind.com/worldwide/support/tech-tips-posters/introduction-to-fetal-bovine-serum-class/
Hope this helps.
Best.
Hi Gabriela Ciapetti
It looks like Malcolm Nobre has given a very detailed answer.
I wonder if a simple solution would be to run the test on untreated cells with and without FBS in the media. The difference between the two ALP values would be due to the FBS, and could then be subtracted from the other results. It may be necessary to repeat this for different batches of FBS as composition is variable by batch.
Good luck!
BWE
I agree with Benjamin W. Elwood, you can do the assay in the presence and absence of FBS and the difference will be due to ALP activity present in FBS.
good luck.
Pls check the Alk p specific activity (Substrate assay PNPP) with cell line extracted for different concentrations from 1 to 10%
Check with the composition of plain media and FBS as well
See that FBS does not inhibit ALP activity
Best wishes
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