Question (50899) | Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369)

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Question

I would like to measure alkaline phosphatase in lysed human vascular smooth cells, using your kit # ab83369. I will be comparing ALP activity values from cells harvested at various time-points; every three days for two weeks. What do recommend for sample preparation and storage for that two week period in order to maintain stability of the enzyme, allowing all samples to be examined in a single assay?

Answer

In this case,washed cells (1×10^5) can be homogenized in the Assay Buffer and centrifuged to remove insoluble material at 13,000g for 3 minutes. The supernatant from this can be aliquoted and stored at -80°C for later analysis. The aliquoting is important because repeated freeze thaw will degrade the enzyme.

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