Overview

  • Product name
    Alkaline Phosphatase Assay Kit (Colorimetric)
    See all Alkaline phosphatase kits
  • Detection method
    Colorimetric
  • Sample type
    Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
  • Assay type
    Enzyme activity (quantitative)
  • Sensitivity
    > 10 µU
  • Range
    10 µU - 250 µU
  • Assay time
    1h 10m
  • Species reactivity
    Reacts with: Other species, Mammals
  • Product overview

    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 other mammalian samples.


    The kit uses p-nitrophenyl phosphate (pNPP) as a phosphatase substrate which turns yellow (λmax= 405 nm) when dephosphorylated by ALP.


    Alkaline phosphatase assay protocol summary:
    - add samples and standards to wells
    - add pNPP solution to sample wells (not to standards)
    - add ALP enzyme solution to standard wells (not to samples)
    - incubate for 60 min at room temp
    - add stop solution
    - analyze with microplate reader

  • Notes

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

  • Platform
    Microplate reader

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components Identifier 500 tests
    ALP Assay buffer NM 1 x 100ml
    ALP Enzyme (Lyophilised) Green 1 vial
    pNPP Substrate (10 tablets) Red 1 vial
    Stop Solution WM 1 x 10ml
  • Research areas
  • Relevance
    Alkaline 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 localization
    Cell 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

Associated products

Images

  • Alkaline phosphatase was measured in Caco-2 cells after 48 hours treatment with PMA and Butyrate using Alkaline Phosphatase assay kit (ab83369).

  • 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.

Protocols

References

This product has been referenced in:
  • Xie L  et al. Potential Biomarkers for Primary Open-Angle Glaucoma Identified by Long Noncoding RNA Profiling in the Aqueous Humor. Am J Pathol 189:739-752 (2019). Read more (PubMed: 30677397) »
  • Morsi NM  et al. Risedronate-Loaded Macroporous Gel Foam Enriched with Nanohydroxyapatite: Preparation, Characterization, and Osteogenic Activity Evaluation Using Saos-2 Cells. AAPS PharmSciTech 20:104 (2019). Read more (PubMed: 30737611) »
See all 114 Publications for this product

Customer reviews and Q&As

31-32 of 32 Abreviews or Q&A

Answer

Please find below the answers provided by the lab where the kit ab83369 is produced :
1. The assay buffer does contain detergents to aid in the lysis of the cells. However, to achieve complete lysis we also recommend that you pass it through the Dounce homogenizer. You can scrape off the cells in the assay biffer and then homogenize the cell sample.
2. ~1X105 cells (in data sheet) should ideally help you to obtain values in the linear range, but this also depends on your samples, so we recommend that you test a couple of dilutions/ volumes of your sample.

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Answer

When using this kit, it is best to determine if the sample you are using has the enzyme or the compound that is being detected in the linear range of the standard graph in order to determine the exact concentration on your sample. Hence we recommend that you test different amounts or dilution of your samples to be in this linear range of the graph. Fresh and frozen sample should work well with the assay (Precaution: in case of frozen samples avoid, repeated freeze thawing).

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31-32 of 32 Abreviews or Q&A

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