製品の概要

  • 製品名
    Alkaline Phosphatase Assay Kit (Colorimetric)
    Alkaline phosphatase キット 製品一覧
  • サンプルの種類
    Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
  • アッセイタイプ
    Quantitative
  • 検出感度
    > 10 µU
  • 検出範囲
    10 µU - 250 µU
  • 種交差性
    交差種: Mammal
  • 製品の概要

    Abcam's 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 biological samples. The kit uses p-nitrophenyl phosphate (pNPP) as a phosphatase substrate which turns yellow (λmax= 405 nm) when dephosphorylated by ALP.
    Visit our FAQs page for tips and troubleshooting.

  • 特記事項

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

  • アプリケーション
    適用あり: Functional Studiesmore details

製品の特性

  • 保存方法
    Store at -20°C. Please refer to protocols.
  • 内容 ラベル 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
  • 研究分野
  • 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
    • 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

アプリケーション

Our Abpromise guarantee covers the use of ab83369 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Functional Studies Use at an assay dependent dilution.

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  • 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.

プロトコール

参考文献

This product has been referenced in:
  • Prajapati MV  et al. Original Research: Evaluation of pulmonary response to inhaled tungsten (IV) oxide nanoparticles in golden Syrian hamsters. Exp Biol Med (Maywood) 242:29-44 (2017). Read more (PubMed: 27534980) »
  • Li Y  et al. Electroactive BaTiO3 nanoparticle-functionalized fibrous scaffolds enhance osteogenic differentiation of mesenchymal stem cells. Int J Nanomedicine 12:4007-4018 (2017). Functional Studies ; Rat . Read more (PubMed: 28603415) »

See all 26 Publications for this product

レビューと Q&A

I confirm this kit can be used with both protein assay kits (BCA and Bradford).


To prepare the samples they should not necessarily require sonication, so I can recommend to follow the protocol.
If there are difficulties lysing the sample, then sonication can be tried. But it is not usually required.

We have not tested the kit with bone samples, but there is no reason for the kit to not work with bone samples. Grinding and homogenization would be critical. The following two references used ab83369 to detect alkaline phosphatase produced in bone tis...

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Unfortunately, the full composition of the buffer is proprietary however, I can tell you that it does contain Brij35 as a detergent to lyse cells but also zinc chloride.

To measure intracellular ALP, washed cells (˜100,000 cells) can be homogenized, preferably using a dounce homogenizer in the Assay Buffer (2-3 times the volume of the cells). Then centrifuge to remove insoluble material at 13,000g for 3 minutes at 4 de...

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10ul of enzyme is used for each well for the standards only. There is 1ml of reconstituted enzyme which is equal to 100 standard wells. Each std has 6 datapoints. With the enzyme mix the standard curve can be run 16 times.
The samples do not need ...

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That note refers to cell culture medium samples, and the control would be a sample of medium that has not been exposed to cells. If your samples are cell lysates, you do not need this control. Your negative control will simply be the Assay Buffer inclu...

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Based on the standard curve equation, y=mx+b, you have to calculate x. This number signifies umoles of pNP for 1 OD. This number is A.

The experimental samples are incubated with pNPP for only 60 minutes before adding stop solution. That is also the case for the incubation of the pNPP standards with the ALP enzyme: 60 minutes of incubation, then stop solution.



Yes, and it can be found in our catalogue as ALP Enzyme (ab134678).

1-10 of 32 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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