Mitochondrial Aldehyde Dehydrogenase (ALDH2) Activity Assay试剂盒(ab115348)


  • 产品名称Mitochondrial Aldehyde Dehydrogenase (ALDH2) Activity Assay试剂盒
    参阅全部 Aldehyde Dehydrogenase 试剂盒
  • 检测方法Colorimetric
  • 精确度
    样品 n Mean SD CV%
    1 20 5%
    样品 n Mean SD CV%
    1 4 12%
  • 测试
    1 x 96 test
  • 样品类型
    Cell culture extracts, Tissue Extracts
  • 检测类型Enzyme activity
  • 范围
    60 µg/ml - 250 µg/ml
  • 实验步骤Multiple steps standard assay
  • 种属反应性
    与反应: Mouse, Rat, Cow, Dog, Human
  • 产品概述

    ab115348 is used to determine mitochondrial aldehyde dehydrogenase activity (ALDH2) in a sample. The enzyme is captured within the wells of the microplate and activity is determined by following the production of NADH in the following ALDH2 catalyzed reaction:

    acetaldehyde + NAD+ -> acid + NADH

    The generation of NADH is coupled to the 1:1 reduction of a reporter dye to yield a colored (yellow) reaction product whose concentration can be monitored by measuring the increase in absorbance at 450 nm (Dye molar extinction coefficient - 37000 M-1cm-1). ab115348 immunocaptures in each well only native ALDH2 from the chosen sample; this removes all other enzymes, including unrelated aldehyde dehydrogenases.

  • 说明

    Reagent dye, Coupler and NAD+ are shipped lyophilized. Before use rehydrate by adding 0.25 mL pure water to each tube and vortex each tube thoroughly to dissolve. After hydration unused amounts of these three materials should be stored at -80°C. Store all other components store at 4°C. This kit is stable for 6 months from receipt.

  • 经测试应用适用于: Functional Studiesmore details
  • 平台Microplate



    Our Abpromise guarantee covers the use of ab115348 in the following tested applications.

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

    应用 Ab评论 说明
    Functional Studies Use at an assay dependent concentration.

    Mitochondrial Aldehyde Dehydrogenase (ALDH2) Activity Assay Kit 图像

    • Reference sample graph
    • Reference Sample graph
    • Determination of effect reactive species on ALDH2 activity.
      The mitochondrial location of ALDH2 may increase the exposure of this enzyme to high levels of reactive oxygen and nitrogen species generated in mitochondria. ALDH2 has been shown to be specifically inactivated by nitrative stress, particularly by modification or crosslinking of an active site cysteine. To demonstrate this rat liver homogenate was exposed in vitro to a dilution series of peroxynitirite at concentrations - 0, 25, 50, 100, 200, 400, 800, 1600 µM. After exposure the samples were extracted, diluted to 0.25 mg/mL and loaded into microplate wells in duplicate. After following the assay protocol as described above, the activity of each sample was measured and graphed against peroxynitrite exposure concentration. The ALDH2 activity after exposure was reduced with an IC50approximately 210 µM.


    Mitochondrial Aldehyde Dehydrogenase (ALDH2) Activity Assay Kit (ab115348)参考文献

    This product has been referenced in:
    • Cummins TD  et al. Metabolic remodeling of white adipose tissue in obesity. Am J Physiol Endocrinol Metab 307:E262-77 (2014). Read more (PubMed: 24918202) »
    • Paneni F  et al. Deletion of the activated protein-1 transcription factor JunD induces oxidative stress and accelerates age-related endothelial dysfunction. Circulation 127:1229-40, e1-21 (2013). Mouse . Read more (PubMed: 23410942) »

    See all 3 Publications for this product

    Product Wall

    Thank you for contacting us.
    I am sorry to hear that you are obtaining low OD readings with the kit.

    Could you please let me know the following details:

    1) What samples were used?
    2) How were they prepared?
    3) What OD...

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    Thank you for contacting us.

    The aldehyde substrate is 25mM final, 1mM b-NAD+ in a buffer at pH9.

    These were all optimized concentrations. To look at competitive inhibitors you could decrease the aldehyde conc...

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    Thank you for contacting us.

    The final pH is 9.0.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Thank you for contacting us.

    This is a species specific phenomenon, most mouse raised monoclonal antibodies have a lower affinity for rodent proteins therefore the activity yield is lower.

    I would predict that the antibody in thi...

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    Is this kit compatible with monkey tissue?
    A: We did not test money sample with this assay. According to UniProtKB, the identity of human ALDH2 and other primate’s are around 98% to 99%. Therefore, this assay most likely will work with monke...

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    Thank you for your enquiry and your interest in our products.

    You need to plate out the cells parallel for the two assays and perform the assays separately. Once you have used the cells for one assay, unfortunately you can't use the same cell...

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    Thank you for your reply.

    I am glad to hear that the problem was solved.

    Please do not hesitate to contact us if you need any more advice or information.

    Thank you for contacting us.

    I have heard back from the lab with the following information:

    All these assays require different sample concentrations described in each booklet protocol - but in most cases is approximately 10ug/micro...

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    Thank you for the extra details.

    I am skeptical that the assay procedure, using acetaldehyde in solution, is the problem. The data you see on the datasheet is generated with the same protocol and reagents.

    My colleague at MitoScie...

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    Yes you are correct it is Dulbecco’s PBS. I hope this is helpful. Please let me know if you need anything further.