製品の概要

  • 製品名

    Total NADP and NADPH Assay Kit (Colorimetric)
    NADP/NADPH キット 製品一覧
  • 検出方法

    Colorimetric
  • サンプルの種類

    Cell culture supernatant
  • アッセイタイプ

    Quantitative
  • 検出感度

    > 0.03 µM
  • 検出範囲

    0.031 µM - 1 µM
  • 全工程の試験時間

    2h 00m
  • 製品の概要

    Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. NADH is the reduced form of NAD+. NAD forms NADP with the addition of a phosphate group to the 2' position of the adenyl nucleotide through an ester linkage. The traditional NAD/NADH and NADP/NADPH assays are based on monitoring the changes in NADH or NADPH absorption at 340 nm. The short UV wavelength of NAD/NADH and NADP/NADPH assays makes the traditional methods suffer low sensitivity and high interference.

    Abcam’s Colorimetric total NADP/NADPH Assay Kit (ab136033) provides a convenient method for detecting total NADP and NADPH. The enzymes in the system specifically recognize NADP/NADPH in an enzyme cycling reaction. There is no need to purify NADP/NADPH from the sample mix. The enzyme cycling reaction significantly increases detection sensitivity. The NADPH probe is a chromogenic sensor that has its maximum absorbance at 460 nm upon NADH reduction. The absorbance maximum increases to ~ 635 nm if the enhancer is added to the assay system. The absorption of the NADPH probe is directly proportional to the concentration of NADPH in the solution. The Colorimetric total NADP and NADPH Assay Kit provides a sensitive assay to detect as little as 0.03 µM total NADP/NADPH in a 100 µL assay volume.

  • 試験プラットフォーム

    Microplate reader

製品の特性

  • 保存方法

    Store at -20°C. Please refer to protocols.
  • 内容 400 tests
    Lysis Buffer 1 x 10ml
    NADP/NADPH Recycling Enzyme Mixture 2 vials
    NADPH Probe 1 x 4ml
    NADPH Probe Buffer 1 x 16ml
    NADPH Standard 1 x 167µg
  • 研究分野

  • 関連性

    NADP (Nicotinamide adenine dinucleotide phosphate) is a coenzyme composed of ribosylnicotinamide 5-phosphate (NMN) coupled by pyrophosphate linkage to the 5-phosphate adenosine 2,5-biphosphate. It serves as an electron carrier in a number of reactions, being alternately oxidised (NADP+) and reduced (NADPH). The oxidative phase of the pentose phosphate pathway is the major source of NADPH in cells, producing approxiamtely 60% of the NADPH required. NADPH provides the reducing equivalents for biosynthetic reactions and the oxidation-reduction involved in protecting against the toxicity of ROS, allowing the regeneration of GSH. NADPH is also used for anabolic pathways, such as lipid synthesis, cholesterol synthesis and fatty acid chain elongation.

画像

  • NADP/NADPH measured in mouse tissue lysates (5-50 mg/mL tested, expressed as quantity per mg of extracted protein; duplicates +/- SD).

  • NADP/NADPH measured cell lysates, where 5e6 cells were lysed in 0.25 mL. RAW samples were diluted 1/25-1/125 times, Jurkat samples were below level of detection (duplicates +/- SD).

  • NADP/NADPH measured in undiluted human biological fluids (duplicates +/- SD).

  • Standard curve with background signal subtracted (duplicates; +/- SD).

  • NADPH dose response was measured with the Colorimetric Total NADP/NADPH Assay Kit in a 96-well white/clear bottom plate using a microplate reader. As low as 0.03 µM of NADPH can be detected with 1 hour incubation (n=3) with absorbance measurement at 460nm.

    The absorbance in blank wells (with the PBS buffer only) is used as a control, and is subtracted from the values for those wells with the NADPH reactions.

     

     

プロトコール

参考文献

This product has been referenced in:

See 1 Publication for this product

レビューと Q&A

There are currently no Customer reviews or Questions for ab186033.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

登録