製品の概要

  • 製品名

    ALDH Activity Assay Kit (Colorimetric)
  • 検出方法

    Colorimetric
  • サンプルの種類

    Cell culture supernatant, Serum, Plasma, Cell Lysate, Tissue Culture Media
  • アッセイタイプ

    Enzyme activity (quantitative)
  • 検出感度

    < 0.5 mU/well
  • 全工程の試験時間

    0h 30m
  • 種交差性

    交差種: Mammals, Other species
  • 製品の概要

    ALDH Activity Assay Kit (Colorimetric) (ab155893) is a simple, fast and reliable method to quantify the ALDH enzymatic activity. In this assay, acetaldehyde is oxidized by ALDH generating NADH which then reduces a colorless probe to a colored product with strong absorbance at 450 nm. The assay can detect < 0.5 mU of ALDH activity in a variety of samples.


    ALDH assay protocol summary:
    - add samples and standards to wells
    - add reaction mix and incubate for 20-60 min
    - analyze with a microplate reader at 5 min and 20-60 min

  • 特記事項

    The NAD-dependent Aldehyde Dehydrogenase (ALDH) plays a vital role in cellular detoxification. It oxidizes various aldehydes and generates the corresponding carboxyolic acid. ALDH have been found in every cellular compartment. Based on its structure and function, ALDH comprises 3 major classes in mammals: Class 1 and Class 3 (the tumor form) are located in the cytosol and include both constitutive and induced forms; Class 2 is located in the mitochondria and only exists as the constitutive form. In humans, the ALDH superfamily consists of 19 genes. The mutation of ALDH genes (loss of function) causes human diseases such as Type II hyperprolinemia, pyridoxine-dependent seizure and hyperammonemia. Recent studies show that increased ALDH activity leads to several types of malignancies, serves as a cancer stem cell marker and correlates with poor prognosis.

  • 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 ラベル 100 tests
    Acetaldehyde Purple 1 x 0.5ml
    ALDH Assay Buffer WM 1 x 25ml
    ALDH Positive Control Green 1 vial
    ALDH Substrate Mix Red 1 vial
    NADH Standard Yellow 1 vial
  • 研究分野

画像

  • Standard curve (60 minutes incubation) showing mean of duplicates (+/- SD) with background reads subtracted.

  • Typical OD of the diffrent positive contols mesuared over 1 hour. 

  • ALDH activity measured in human biological fluids (T1=5 min; T2=60 min), showing activity (mU) per mL of tested sample, background signal subtracted.

プロトコール

参考文献

This product has been referenced in:

  • Ouyang S  et al. LncRNA BCAR4, targeting to miR-665/STAT3 signaling, maintains cancer stem cells stemness and promotes tumorigenicity in colorectal cancer. Cancer Cell Int 19:72 (2019). Read more (PubMed: 30962766) »
  • Norona LM  et al. Editor's Highlight: Modeling Compound-Induced Fibrogenesis In Vitro Using Three-Dimensional Bioprinted Human Liver Tissues. Toxicol Sci 154:354-367 (2016). Functional Studies . Read more (PubMed: 27605418) »
See all 2 Publications for this product

レビューと Q&A

1-4 of 4 Abreviews or Q&A

Measurement of ALDH activity in cancer cells

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Detailed protocol was easy followed as per kit manual for measuring the ALDH activity in lung cancer cell lines. The data is consistent with the expression level of ALDH protein in the respective cancer cell lines.

Abcam user community

Verified customer

投稿 Aug 21 2018

Answer

The ALDH Activity Assay Kit (Colorimetric) (ab155893) has not been tested yet in cancer cell lines such as MDA-MB-231 and MCF-7.
Unfortunately, we do not currently have more information about this product. We will update the on-line datasheet for this product as soon as we receive additional data.

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Answer

We would recommend using our assay buffer for the lysis, as RIPA may not contain all the components required for the reaction to occur. The samples should be homogenized in ice cold assay buffer for 10 minutes on ice (page 8 of the online protocol). Lysates prepared in the assay buffer can also be used in Western blot and ELISA.

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