Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369)
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
Overview
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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: Mammals, Other species -
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 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 -
Notes
This kit contains 10 substrate tablets providing convenience for multiple usages.
pNPP Assays
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.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components Identifier 500 tests ALP Assay buffer NM 1 x 100ml ALP Enzyme (Lyophilised) White 1 vial pNPP Substrate (10 tablets) Red 1 vial Stop Solution WM 1 x 10ml -
Research areas
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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
see all
Images
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ALP activity in cells undergoing osteogenic differentiation Image courtesy of Hermenean A et al. Sci Rep. 2017; 7: 16641. doi: 10.1038/s41598-017-16599-5
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.
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Effects of mineralocorticoid receptor blockade on phosphate-induced vascular calcification in vitro Image courtesy of Alesutan I et al. Sci Rep. 2017; 7: 2059. doi: 10.1038/s41598-017-01882-2. Reproduced under the creative commons license http://creativecommons.org/licenses/by/4.0/.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).
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Alkaline phosphatase activity measured in the study of osteoporosis Image from Lee YM. et al., Biomed Res Int. 2019 Feb 19;2019:5073085. Fig 6b doi: 10.1155/2019/5073085. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/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.
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Functional Studies - Alkaline Phosphatase Colorimetric Assay Kit (ab83369) Image from Nepelska M et al., PLoS One 7(12), fig 8c. Doi: 10.1371/journal.pone.0052869. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/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).
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Functional Studies - Alkaline Phosphatase Colorimetric Assay Kit (ab83369)Standard curve: mean of duplicates (+/- SD) with background reads subtracted
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Functional Studies - Alkaline Phosphatase Colorimetric Assay Kit (ab83369)ALP measured in cell culture supernatants showing activity (U) per L of tested sample. Samples were diluted 1-16 fold.
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Functional Studies - Alkaline Phosphatase Colorimetric Assay Kit (ab83369)ALP measured in biological fluids showing activity (U) per L of tested sample. Samples were diluted 4-64 fold.
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Functional Studies - Alkaline Phosphatase Colorimetric Assay Kit (ab83369)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.
