製品の概要

  • 製品名
  • サンプルの種類
    Adherent cells, Suspension cells
  • アッセイタイプ
    Enzyme activity
  • 全工程の試験時間
    1h 00m
  • 製品の概要

    For a more sensitive LDH Assay Kit to use as a cytotoxicity assay, we recommend ab65393.


    LDH Assay Kit (Cytotoxicity) ab65391 provides a fast and simple method for quantitating cytotoxicity based on the measurement of activity of lactate dehydrogenase (LDH) released from damaged cells.


    The assay takes ~0.5-1 hr. LDH activity can be determined by a coupled enzymatic reaction: LDH oxidizes lactate to pyruvate which then reacts with tetrazolium salt INT to form formazan. The increase in the amount of formazan produced in culture supernatant directly correlates to the increase in the number of lysed cells. The formazan dye is water-soluble and can be detected by spectrophotometer at 500 nm.


    This kit was previously called LDH-Cytotoxicity Assay Kit I.

  • 特記事項

    Cell death or cytotoxicity is classically evaluated by the quantification of plasma membrane damage. Unlike many other cytoplasmic enzymes which exist in many cells either in low amount (e.g., alkaline and acid phosphatase) or unstable, LDH is a stable cytoplasmic enzyme present in all cells and rapidly released into the cell culture supernatant upon damage of the plasma membrane.

    If you would like to use a fluorometric reading, please refer to LDH-Cytotoxicity Assay Kit (Fluorometric) (ab197004).

  • 試験プラットフォーム
    Microplate reader

製品の特性

画像

  • LDH release measured at various cell amounts

     

  • Jurkat cells were cultured in a 96-well plate in 100 µl of culture medium. LDH assay was performed using 10 µl of culture medium according to the kit instructions. Light bar: Low control; Dark bar: High control.

プロトコール

参考文献

This product has been referenced in:
  • Gouriou Y  et al. Mitochondrial Ca2+ uptake from plasma membrane Cav3.2 protein channels contributes to ischemic toxicity in PC12 cells. J Biol Chem 288:12459-68 (2013). Read more (PubMed: 23508951) »
  • Spampanato C  et al. Transcription factor EB (TFEB) is a new therapeutic target for Pompe disease. EMBO Mol Med 5:691-706 (2013). Read more (PubMed: 23606558) »
See all 2 Publications for this product

レビューと Q&A

1-9 of 9 Abreviews or Q&A

Answer

Thank you for contacting us.

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

There are numerous differences between these two assay kits. For details, I have attached the datasheets, but here are the main differences:






Kit
ab65391
ab65393

Detection wavelength
500 nm
450 nm

Principle
LDH activity can be determined by a
coupled enzymatic reaction: LDH oxidizes lactate to pyruvate which then reacts with
tetrazolium salt INT to form formazan. The increase in the amount of formazan produced in
culture supernatant directly correlates to the increase in the number of lysed cells.
The assay utilizing an enzymatic coupling reaction: LDH oxidizes
lactate to generate NADH, which then reacts with WST to generate yellow color. The intensity
of the generated color correlates directly with the cell number lysed.

Special advantages:

Since WST is brighter,
less amount of culture medium is required for the assay, and thus the background from serum
and culture medium is significantly reduced. Using the assay, cells can be cultured in regular
10% serum containing medium, no reducing serum or special medium is required for the
assay. In addition, since the WST is more stable, the reaction can be read multiple times, and
can also be stopped at any time point during the reaction. LDH activity can be easily quantified
by spectrophotometer or plate reader at OD450 nm. The kit provides all necessary reagents
including LDH positive control.






One can use only the cell media with these kits.


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

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Answer

Thank you for your reply, and I'm sorry that we could not solve this problem for you.

I'm happy to issue the refund. Do you have your order number, PO number, or date of the purchase?

Please let me know if you have any questions or if there is anything else that we can do for you.

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Answer

Thank you for your call this week and for your patience while I have been in touch with the lab.

The scientists at the lab have confirmed that the absorbance value should increase with increasing cytotoxicity, so the data is contradicting what we would expect to see.

We do have a couple of additional questions:

1) 1) Is the control media incubated in the same wayas the other samples?
2) 2) Which kind of microtiter plate areyou using?
3) 3) What wavelengthis usedto measure absorbance?
4) 4) Is it possible that the PLX treated samples were mislabelled as the media standards and vice versa?
I appreciate your patience and I look forward to hearing from you. Please note that this kit is covered by our Abpromise guarantee, so if you prefer a replacement, credit, or refund I will be happy to set that up for you.
Please let me know if you have any questions or if there is anything else that we can do for you.

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Answer

Thank you for contacting us and your interest in our products.

The LDH-Cytotoxicity Assay Kit (ab65391) is designed to measure the levels of LDH in the cell culture supernatants and determines the cytotoxicitybased on this. The kit can be theoretically be optimized by the end user for tissue samples, by testing the growth media for LDH or maybe using the tissue lysate withthe kit. However since we have not performed any optimization for tissue samples this would need to be ascertained by the end user.

I hope this information has been of help. If you require any further assistance please do not hesitate to contact us again.

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Answer

Thank you for contacting us. This kit is compatible with mammalian samples and hence will work with mouse and rat samples. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.    

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Answer

Thank you for your inquiry. There are a few differences between those two kits: With the LDH-Cytotoxicity Assay Kit ab65391 LDH activity is determined by a coupled enzymatic reaction. This assay kit is sensitive, convenient, and precise, and is applicable to a variety of cytotoxicity studies. Reagents are provided for 400 assay tests. The LDH-Cytotoxicity Assay Kit II ab65393 utilizes an advanced reagent (WST) for a fast and more sensitive detection of LDH released from damaged cells. Since WST is brighter, less culture medium is required for the assay, and thus the background from serum and culture medium is significantly reduced. Using the assay, cells can be cultured in regular 10% serum containing medium, no reducing serum or special medium is required for the assay. In addition, since the WST is more stable, the reaction can be read multiple times, and can also be stopped at any time point during the reaction. Furthermore, this kit contains the reagents for 500 assays to be run. I hope this information is helpful to you. Please do not hesitate to contact me if you need any further advice or information in this regard.

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Question
Answer

Merci de nous avoir contactés. Mes collaborateurs m'ont informé que le kit ab102526 est le plus adapté à vos besoins. Le lien vers la fiche technique est : www.abcam.com/ab102526. J'espère que cette information vous est utile. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.      

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Answer

As long as your plate reader has the ability to measure the absorbance of all samples at 490-500 nm, then you will be able to use the plate reader with the LDH-Cytotoxicity assay kit. I believe that the plate reader that you have, does have the ability to read wavelengths at 490-500nm but I would consult with the user guide for the machine to confirm this.

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Question
Answer

Thank you for your phone call today and for your questions about our LDH assay kits. The kits do utilize different reactions in order to determine DLH activity, but there are also some other differences. I have summarized the advantages of each kit below. Ab65391: Sensitivity of 10,000 cells. Fast and simple method for quantifying cytotoxicity. Applicable to a variety of cytotoxicity studies. Ab65393: Sensitivity of 20,000 cells. Uses WST so less culture medium is required. Background from serum and medium is reduced. WST is more stable, so the reaction can be read multiple times and can be stopped at any time. Cells can be cultured in regular medium containing 10% serum. I hope this information is helpful, but please let me know if you have any further questions or if there is anything else that we can do for you.

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

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