• Product nameAlkaline Phosphatase Assay Kit (Colorimetric)
    See all Alkaline phosphatase kits
  • Tests
    500 x 1 assay
  • Sample type
    Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
  • Assay typeQuantitative
  • Sensitivity
    > 10 µU
  • Range
    10 µU - 250 µU
  • Species reactivity
    Reacts with: A wide range of mammals
  • Product overview

    Abcam's Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369) is a highly sensitive, simple, direct and HTS-ready colorimetric assay designed to measure alkaline phosphatase (ALP) activity in serum and biological samples. The kit uses p-nitrophenyl phosphate (pNPP) as a phosphatase substrate which turns yellow (λmax= 405 nm) when dephosphorylated by ALP.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    This kit contains 10 substrate tablets providing convenience for multiple usages.

  • Tested applicationsFunctional Studiesmore details


  • RelevanceAlkaline phosphatase (ALP) catalyzes the hydrolysis of phosphate esters in alkaline buffer and produces an organic radical and inorganic phosphate. Changes in alkaline phosphatase level and activity are associated with various disease states in the liver and bone.
  • Cellular localizationCell membrane; Lipid-anchor, GPI-anchor.
  • Alternative names
    • Alkaline phosphatase intestinal
    • Alkaline phosphatase liver/bone/kidney
    • Alkaline phosphatase liver/bone/kidney isozyme
    • Alkaline phosphatase tissue nonspecific isozyme
    • Alkaline phosphomonoesterase
    • ALPI
    • ALPL
    • AP TNAP
    • Glycerophosphatase
    • HOPS
    • IAP
    • Intestinal alkaline phosphatase
    • Kasahara isozyme
    • Liver/bone/kidney type alkaline phosphatase
    • Tissue nonspecific ALP
    • TNAP
    • TNSALP
    see all


Our Abpromise guarantee covers the use of ab83369 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Functional Studies Use at an assay dependent dilution.

Alkaline Phosphatase Assay Kit (Colorimetric) images

  • Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • ALP measured in cell culture supernatants showing activity (U) per L of tested sample. Samples were diluted 1-16 fold.

  • ALP measured in biological fluids showing activity (U) per L of tested sample. Samples were diluted 4-64 fold.

  • Measurement of ALP activity using ab83369. a. ALP activity in fresh medium (80 μL, without culturing), 3 day old HeLa cell culture medium (80 μL) and human serum (80 μL, 1/10 dilution). b. ALP activity in HeLa cells. 5 x 104 HeLa cells were homogenized in 1 mL of Assay Buffer, diluted 1/10 in Assay Buffer, and 80 μL used for the measurement.


References for Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369)

This product has been referenced in:
  • Wessel Stratford E  et al. The tankyrase-specific inhibitor JW74 affects cell cycle progression and induces apoptosis and differentiation in osteosarcoma cell lines. Cancer Med 3:36-46 (2014). Functional Studies . Read more (PubMed: 24403055) »
  • Hu J  et al. Filamin B regulates chondrocyte proliferation and differentiation through Cdk1 signaling. PLoS One 9:e89352 (2014). Functional Studies . Read more (PubMed: 24551245) »

See all 14 Publications for this product

Product Wall

I confirm this kit can be used with both protein assay kits (BCA and Bradford).

To prepare the samples they should not necessarily require sonication, so I can recommend to follow the protocol.
If there are difficulties lysing the sample, then sonication can be tried. But it is not usually required.

We have not tested the kit with bone samples, but there is no reason for the kit to not work with bone samples. Grinding and homogenization would be critical. The following two references used ab83369 to detect alkaline phosphatase produced in bone tis...

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Unfortunately, the full composition of the buffer is proprietary however, I can tell you that it does contain Brij35 as a detergent to lyse cells but also zinc chloride.

To measure intracellular ALP, washed cells (˜100,000 cells) can be homogenized, preferably using a dounce homogenizer in the Assay Buffer (2-3 times the volume of the cells). Then centrifuge to remove insoluble material at 13,000g for 3 minutes at 4 de...

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10ul of enzyme is used for each well for the standards only. There is 1ml of reconstituted enzyme which is equal to 100 standard wells. Each std has 6 datapoints. With the enzyme mix the standard curve can be run 16 times.
The samples do not need ...

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That note refers to cell culture medium samples, and the control would be a sample of medium that has not been exposed to cells. If your samples are cell lysates, you do not need this control. Your negative control will simply be the Assay Buffer inclu...

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Based on the standard curve equation, y=mx+b, you have to calculate x. This number signifies umoles of pNP for 1 OD. This number is A.

The experimental samples are incubated with pNPP for only 60 minutes before adding stop solution. That is also the case for the incubation of the pNPP standards with the ALP enzyme: 60 minutes of incubation, then stop solution.

Yes, and it can be found in our catalogue as ALP Enzyme (ab134678).