Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 1 hr
- Sample type: Cell culture media, Cell Lysate, Other biological fluids, Plasma, Saliva, Serum, Tissue Extracts, Urine
Product nameAlkaline Phosphatase Assay Kit (Fluorometric)
See all Alkaline phosphatase kits
Sample typeSaliva, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
Assay typeEnzyme activity (quantitative)
Assay time1h 00m
Species reactivityReacts with: Mammals, Other species
Alkaline Phosphatase Assay Kit (Fluorometric) (ab83371) is a highly sensitive kit to detect alkaline phosphatase (ALP) activity in serum and other biological samples. In the alkaline phosphatase assay protocol, ALP cleaves the phosphate group of the non-fluorescent 4-Methylumbelliferyl phosphate disodium salt (MUP) substrate, resulting in an increased fluorescent signal (Ex/Em = 360nm/440nm) when the substrate is dephosphorylated.
The kit is an ultra sensitive, simple, direct and HTS-ready assay designed to measure ALP activity with a detection sensitivity ~1 µU, more sensitive than colorimetric assays.
Alkaline phosphatase assay protocol summary:
- add samples and standards to wells
- add reaction mix to wells
- add ALP enzyme solution to standard wells (not to samples)
- incubate for 30 min at room temp
- add stop solution
- analyze with microplate reader
Alkaline phosphatase (ALP) catalyzes the hydrolysis of phosphate esters in alkaline buffer and produces an organic radical and inorganic phosphate. The change in alkaline phosphatase level and activity is associated with many diseases in the liver and bones.
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 500 tests ALP Assay Buffer NM 1 x 100ml ALP Enzyme (Lyophilised) White 1 vial MUP Substrate Red 1 vial Stop Solution WM 1 x 25ml
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
Effects of IL-17A and/or TNF-α on alkaline phosphatase activity are shown. hMSCs were plated at a density of 5 x 103 cell/cm2, cells were cultured in osteogenic medium for 14 days, in the absence (0) or presence of TNF-α 1 ng/ml and/or IL-17A 50 ng/ml. ALP activity was measured by fluorometry useing ab83371. Results were analyzed using the Wilcoxon test. ∗p < 0.05; ∗∗p < 0.005 vs. induction medium alone.
Alkaline phosphatase measured in mouse tissue lysates showing activity (mU) per mg protein of tested sample
Alkaline phosphatase measured in biological fluids showing quantity (mU) per microliter of tested sample
4-Methylumbelliferone (4-MU) standard curve.
ab83371 has been referenced in 22 publications.
- Fernandes BF et al. Laser Nd:YAG patterning enhance human osteoblast behavior on zirconia implants. Lasers Med Sci 35:2039-2048 (2020). PubMed: 32556830
- Li SJ et al. HDAC I inhibitor regulates RUNX2 transactivation through canonical and non-canonical Wnt signaling in aortic valvular interstitial cells. Am J Transl Res 11:744-754 (2019). PubMed: 30899376
- Amer MH et al. A biomaterials approach to influence stem cell fate in injectable cell-based therapies. Stem Cell Res Ther 9:39 (2018). PubMed: 29467014
- Xia Y et al. Gold nanoparticles in injectable calcium phosphate cement enhance osteogenic differentiation of human dental pulp stem cells. Nanomedicine 14:35-45 (2018). PubMed: 28887211
- Wan Hasan WN et al. Annatto-derived tocotrienol stimulates osteogenic activity in preosteoblastic MC3T3-E1 cells: a temporal sequential study. Drug Des Devel Ther 12:1715-1726 (2018). PubMed: 29942115