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
- Sensitivity: 10 µU
製品の概要
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製品名
Alkaline Phosphatase Assay Kit (Colorimetric)
Alkaline phosphatase キット 製品一覧 -
検出方法
Colorimetric -
サンプルの種類
Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media -
アッセイタイプ
Enzyme activity (quantitative) -
検出感度
> 10 µU -
検出範囲
10 µU - 250 µU -
全工程の試験時間
1h 10m -
種交差性
交差種: Mammals, Other species -
製品の概要
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.
How the assay works
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 as part of an ALP activity assay. This kit contains 10 substrate tablets providing convenience for multiple usages.
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
Related alkaline phosphatase assay products
For a fluorometric alkaline phosphatase assay kit, we recommend ab83371. This assay is based on the 4-Methylumbelliferyl phosphate disodium salt (MUP) substrate. Alkaline phosphatase cleaves the phosphate group of this substrate, releasing fluorescent 4-methylumbelliferyl (Ex/Em = 360nm/440nm).
For a luminescent alkaline phosphatase assay, we recommend ab233466. This assay is bsed on the ability of alkaline phosphatase to remove the phosphate group from D-luciferin phosphate, releasing the luciferin to be processed by luciferase.
For alkaline phosphatase staining, we recommend Alkaline Phosphatase Staining Kit ab284936. This kit is based on the substrate, 5-Bromo-4-chloro-3-indolyl phosphate (BCIP) which is processed by alkaline phosphatase to form a blue colored intermediate. Nitro Blue Tetrazolium (NBT) then oxidizes the intermediate to form an insoluble, dark blue dimer, which can be viewed by microscopy.
Getting the best performance from ab83369
Technical Note: Alkaline phosphatase is unlikely to be detectable in healthy urine but may be elevated in certain kidney, liver or bone disorders.
Alkaline phosphatase assay methods
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.
The pNPP phosphatase assay method is by far the most popular method for a colorimetric alkaline phosphatase assay for liquid samples or cell/tissue lysates.
Other notes
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. -
特記事項
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the SDS download section.
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試験プラットフォーム
Microplate reader
製品の特性
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保存方法
Store at -20°C. Please refer to protocols. -
内容 500 tests 10000 tests ALP Assay Buffer I 1 x 100ml 20 x 100ml Enzyme Mix II 1 vial 20 vials pNPP Substrate 1 vial 20 vials Stop Solution I 1 x 10ml 20 x 10ml -
研究分野
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関連性
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. -
細胞内局在
Cell membrane; Lipid-anchor, GPI-anchor. -
別名
- Alkaline phosphatase intestinal
- Alkaline phosphatase liver/bone/kidney
- Alkaline phosphatase liver/bone/kidney isozyme
see all
関連製品
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Related Products
画像
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ALP activity in cells undergoing osteogenic differentiationImage 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 vitroImage 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 osteoporosisImage 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.
The serum levels of alkaline phosphatase (ALP) were measured using assay kits (ab83369, Alkaline Phosphatase Assay Kit; Abcam) according to the manufacturer's instructions. Briefly, 80 μL of sample dilutions, 50 μL of 5 mM p-nitrophenyl phosphate (pNPP) solution, and 10 μL of ALP enzyme solution were added sequentially to the reaction wells and the plate was incubated at 25°C for 60 min protected from light. The reaction was halted by adding 20 μL stop solution, and then the absorbance was measured at 405 nm. ALP activity was calculated using a standard curve.
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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.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (307)
ab83369 は 307 報の論文で使用されています。
- Pugazhendhi AS et al. Extracellular Proteins Isolated from L. acidophilus as an Osteomicrobiological Therapeutic Agent to Reduce Pathogenic Biofilm Formation, Regulate Chronic Inflammation, and Augment Bone Formation In Vitro. Adv Healthc Mater 13:e2302835 (2024). PubMed: 38117082
- Erdogan YK & Ercan B Anodized Nanostructured 316L Stainless Steel Enhances Osteoblast Functions and Exhibits Anti-Fouling Properties. ACS Biomater Sci Eng 9:693-704 (2023). PubMed: 36692948
- Shosha HM et al. Effect of monosodium glutamate on fetal development and progesterone level in pregnant Wistar Albino rats. Environ Sci Pollut Res Int 30:49779-49797 (2023). PubMed: 36787072
- D'Amico E et al. Emerging Effects of Resveratrol Derivatives in Cells Involved in Oral Wound Healing: A Preliminary Study. Int J Mol Sci 24:N/A (2023). PubMed: 36834684
- Felice P et al. Osteoblasts and Fibroblasts Interaction with a Porcine Acellular Dermal Matrix Membrane. Int J Mol Sci 24:N/A (2023). PubMed: 36835067