Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 1 hr 10 min
- Sample type: Cell culture media, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine
- Sensitivity: 10 µU
Product nameAlkaline Phosphatase Assay Kit (Colorimetric)
See all Alkaline phosphatase kits
Sample typeUrine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
Assay typeEnzyme activity (quantitative)
Sensitivity> 10 µU
Range10 µU - 250 µU
Assay time1h 10m
Species reactivityReacts with: Mammals, Other species
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 is a pNPP assay; it 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
This product is manufactured by BioVision, an Abcam company and was previously called K412 Alkaline Phosphatase Activity Colorimetric Assay Kit. K412-500 is the same size as the 500 test size of ab83369.
This kit contains 10 substrate tablets providing convenience for multiple usages.
pNPP assays are a class of phosphatase assays using p-nitrophenyl phosphate (pNPP) as a phosphatase substrate. They can be used to measure alkaline phosphatase, neutral phosphatase, and acid phosphatase activity. These different types of phosphatases are active respectively in alkaline, neutral, and acid assay buffers. pNPP assays include Alkaline Phosphatase Assay Kit ab83369 and Acid Phosphatase Assay Kit ab83367.
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 500 tests 10000 tests ALP Assay Buffer I NM 1 x 100ml 20 x 100ml Enzyme Mix II White 1 vial 20 vials pNPP Substrate Red 1 vial 20 vials Stop Solution I WM 1 x 10ml 20 x 10ml
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.
- Alkaline phosphatase intestinal
- Alkaline phosphatase liver/bone/kidney
- Alkaline phosphatase liver/bone/kidney isozyme
Hermenean A et al. (2017) measured alkaline phosphatase activity in cells cultured in CHT/GO (3D chitosan scaffolds enriched with graphene oxide) biomaterials for up to 28 days using ab83369.
Alesutan I et al. (2017) measured alkaline phosphatase activity in vitro in primary human aortic smooth muscle cells using ab83369. Cells were treated with phosphate (pi) or without (Ctr, and with or without additional treatment of spironolactone (Spr) or 10 µM eplerenone (Epl).
Lee YM et al. (2019) used alkaline phosphatase assay kit ab83369 to measure the serum levels of alkaline phosphatase as a marker of bone turnover. Rat groups include sham-operated (Sham), OVX-control (OVX), E2-treated OVX groups (E2), and LABE 500 mg/kg treated OVX groups (LABE) (N=10/group).
Serum ALP levels in the OVX group were significantly increased compared to those in Sham group, and oral administration of LABE (500 mg/kg) significantly reduced the serum ALP levels as an osteoporosis-related serum marker.
Nepelska M et al. (2012) assayed alkaline phosphatase activity 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.
ab83369 has been referenced in 273 publications.
- Haseli M et al. Crosslinked Layered Surfaces of Heparin and Poly(L-Lysine) Enhance Mesenchymal Stromal Cell Behavior in the Presence of Soluble Interferon Gamma. Cells Tissues Organs 212:8-20 (2023). PubMed: 34937023
- Jia CQ et al. A biomimetic gradient porous cage with a micro-structure for enhancing mechanical properties and accelerating osseointegration in spinal fusion. Bioact Mater 23:234-246 (2023). PubMed: 36439084
- Shi H et al. MicroRNAs in Serum Exosomes as Circulating Biomarkers for Postmenopausal Osteoporosis. Front Endocrinol (Lausanne) 13:819056 (2022). PubMed: 35360081
- Kotlarz M et al. Bioprinting of Cell-Laden Hydrogels onto Titanium Alloy Surfaces to Produce a Bioactive Interface. Macromol Biosci 22:e2200071 (2022). PubMed: 35365963
- Fakhr Y et al. Tumor Necrosis Factor-α Induces a Preeclamptic-like Phenotype in Placental Villi via Sphingosine Kinase 1 Activation. Int J Mol Sci 23:N/A (2022). PubMed: 35409108