Overview

  • Product nameLDH-Cytotoxicity Assay Kit II (500 assays)
    See all Lactate dehydrogenase kits
  • Tests
    500 x 1 assay
  • Sample type
    Adherent cells, Suspension cells
  • Assay typeQuantitative
  • Assay time
    1h 00m
  • Product overview

    Abcam's LDH-Cytotoxicity Assay Kit II utilizes the advanced WST reagent for a fast and more sensitive detection of LDH released from damaged 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. 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. 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 OD450nm. The kit provides all necessary reagents including LDH positive control.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    Cell death or cytotoxicity is classically evaluated by the quantification of plasma membrane damage. Lactate dehydragenase (LDH) is a stable enzyme, present in all cell types, and rapidly released into the cell culture medium upon damage of the plasma membrane. LDH, therefore, is the most widely used marker in cytotoxicity study.

    If you need to purchase additional Cell Lysis Solution for this product, please see ab128687.

  • Tested applicationsFunctional Studiesmore details

Properties

  • RelevanceLactate dehydrogenase (LDH) is an oxidoreductase which catalyses the interconversion of pyruvate and lactate with concomitant interconversion of NADH and NAD+. As it can also catalyze the oxidation of hydroxybutyrate, it is occasionally called Hydroxybutyrate Dehydrogenase (HBD). There are 5 different isoenzymes of LDH, LDH1 to LDH5, each composed of 4 subunits which may be of 2 different types - M and H subunits. These subunits are encoded by two different genes: The M subunit is encoded by gene LDHA whilst the H subunit is encoded by LDHB. Usually LDH2 is the predominant form in the serum. An LDH1 level higher than the LDH2 level suggests myocardial infarction (damage to heart tissues releases heart LDH, which is rich in LDH1, into the bloodstream).
  • Cellular localizationCytoplasmic
  • Alternative names
    • L lactate dehydrogenase A chain
    • L lactate dehydrogenase B chain
    • Lactate dehydrogenase A
    • Lactate dehydrogenase B
    • LDHA
    • LDHB
    • Renal carcinoma antigen NY REN 46
    • Renal carcinoma antigen NY REN 59
    see all

Applications

Our Abpromise guarantee covers the use of ab65393 in the following tested applications.

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

Application Abreviews Notes
Functional Studies Use at an assay dependent dilution.

LDH-Cytotoxicity Assay Kit II (500 assays) images

  • Comparison of WST-1 and INT based assays. 3T3 cells were cultured in a 96-well plate in 100 µl of culture medium. LDH assays were performed using 10 µl of culture medium using WST-1 (Brown bar) and INT (Green bar) methods. The WST-1 based assay is more stable and sensitive than the INT based method.

Protocols

References for LDH-Cytotoxicity Assay Kit II (500 assays) (ab65393)

This product has been referenced in:
  • Chaturvedi M  et al. Tissue inhibitor of matrix metalloproteinases-1 loaded poly(lactic-co-glycolic acid) nanoparticles for delivery across the blood-brain barrier. Int J Nanomedicine 9:575-88 (2014). Read more (PubMed: 24531257) »
  • Poon IKh  et al. Phosphoinositide-mediated oligomerization of a defensin induces cell lysis. Elife 3:e01808 (2014). Read more (PubMed: 24692446) »

See all 9 Publications for this product

Product Wall

The kit ab102526 is more suitable for measuring the LDH activity in tissue culture supernatants - It can also be used for tissue extracts.

ab65393 is only tested with cells and cell extracts - tissue extracts are yet to be tested therefore I ...

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Yes, the lab suggested adding 100ul of the media to be tested to 100ul of the LDH reaction mix then read optically. You can make/use a standard curve if you know the conc of the LDH you use for it. We have not provided the conc of LDH in our kit due...

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You will be using a 96 well plate for this assay, and each well has a capacity for 200 µl of the sample. If you take 10 µl of the media, your final volume in each well is only 110 µl. therefore in effect you can use 100 ul of the samp...

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Thank you for your inquiry and your patience with this reply.

I've asked the supplying lab for more information about the components of the stop solution, but they were unable to disclose what they use since they consider it "proprietary...

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The cell lysis solution should be used to prepare the High control only. Please use 10% of cell lysis solution i.e. for 100 ul use 10 ul cell lysis solution.

I hope this information is helpful to you. Plea...

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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: Read More

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Can you please clarify the difference between your CON samples and your LC? It seems like both sets of samples are cells without treatment, is that correct?

What sort of background readings were you gett...

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This kit is compatible with cells. Therefore I just want to make sure that you are planning on using this assay with the BAL cells as well.

In that case you may have to incubate the BAL fluid with the ce...

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We haven't tested this kit with plant material yet so I am sorry we are not aware of any interference of photochemicals with the absorbance reaction of this kit. This kit uses WST1 compound which has been successfu...

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Thank you very much for letting me know. I am pleased to know the kit is now working well for you.
If you have any questions or concerns in the future, please do not hesitate to contact us again.
Until then, I wish you all the best with your ...

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