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    products/assay-kits/ldh-assay-kitcytotoxicity-ab65391.pdf

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LDH Assay Kit (Cytotoxicity) (ab65391)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (9)References (30)

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Functional studies - ab65391 LDH-Cytotoxicity Assay Kit (400 assays)
  • Functional Studies - Lactate Dehydrogenase-Cytotoxicity Assay Kit (ab65391)

Key features and details

  • Assay type: Enzyme activity
  • Platform: Microplate reader
  • Assay time: 1 hr
  • Sample type: Adherent cells, Suspension cells

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Overview

  • Product name

    LDH Assay Kit (Cytotoxicity)
    See all Lactate Dehydrogenase kits
  • Sample type

    Adherent cells, Suspension cells
  • Assay type

    Enzyme activity
  • Assay time

    1h 00m
  • Product overview

    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.

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called K311 LDH-Cytotoxicity Colorimetric Assay Kit. K311-400 is the same size as the 400 test size of ab65391.

    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).

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 400 tests 8000 tests
    Catalyst (Lyophilized) 1 vial 20 vials
    Dye Solution 1 x 45ml 20 x 45ml
  • Research areas

    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Cancer
    • Tumor biomarkers
    • Enzymes
    • Lactic Dehydrogenase
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Sugar Assays
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Cell viability, plasma membrane damage
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Cell Damage
  • Pathway

    Fermentation; pyruvate fermentation to lactate; (S)-lactate from pyruvate: step 1/1.
  • Involvement in disease

    Defects in LDHA are the cause of glycogen storage disease type 11 (GSD11) [MIM:612933]. A metabolic disorder that results in exertional myoglobinuria, pain, cramps and easy fatigue.
  • Sequence similarities

    Belongs to the LDH/MDH superfamily. LDH family.
  • Post-translational
    modifications

    ISGylated.
  • Cellular localization

    Cytoplasm.
  • Target information above from: UniProt accession P00338 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • Cell proliferation-inducing gene 19 protein
    • GSD11
    • L lactate dehydrogenase B chain
    • L-lactate dehydrogenase A chain
    • Lactate dehydrogenase A
    • Lactate dehydrogenase B
    • Lactate dehydrogenase c variant 1
    • Lactate dehydrogenase c variant 3
    • Lactate dehydrogenase c variant 4
    • Lactate dehydrogenase C4
    • Lactate dehydrogenase H chain
    • Lactate dehydrogenase M
    • LDH A
    • LDH B
    • LDH H
    • LDH heart subunit
    • LDH M
    • LDH muscle subunit
    • LDH-A
    • LDH-M
    • LDH1
    • ldha
    • LDHA_HUMAN
    • LDHBD
    • LDHM
    • MS1111
    • PIG19
    • Proliferation inducing gene 19
    • Proliferation-inducing gene 19
    • Renal carcinoma antigen NY REN 46
    • Renal carcinoma antigen NY-REN-59
    • TRG 5
    • TRG5
    see all

Associated products

  • Assay kits

    • LDH Assay Kit / Lactate Dehydrogenase Assay Kit (Colorimetric) (ab102526)
    • Lactate Dehydrogenase (LDH) Assay Kit (Fluorometric) (ab197000)
    • LDH-Cytotoxicity Assay Kit (Fluorometric) (ab197004)
    • LDH Assay Kit (Cytotoxicity) (ab65393)

Images

  • Functional studies - ab65391 LDH-Cytotoxicity Assay Kit (400 assays)
    Functional studies - ab65391 LDH-Cytotoxicity Assay Kit (400 assays)

    LDH release measured at various cell amounts

     

  • Functional Studies - Lactate Dehydrogenase-Cytotoxicity Assay Kit (ab65391)
    Functional Studies - Lactate Dehydrogenase-Cytotoxicity Assay Kit (ab65391)
    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.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (30)

Publishing research using ab65391? Please let us know so that we can cite the reference in this datasheet.

ab65391 has been referenced in 30 publications.

  • Li Y  et al. Scutellaria barbata Inhibits Hepatocellular Carcinoma Tumorigenicity by Inducing Ferroptosis of Hepatocellular Carcinoma Cells. Front Oncol 12:693395 (2022). PubMed: 35321425
  • Borkar NA  et al. Kisspeptins inhibit human airway smooth muscle proliferation. JCI Insight 7:N/A (2022). PubMed: 35420998
  • Tolardo V  et al. In Vitro High-Throughput Toxicological Assessment of Nanoplastics. Nanomaterials (Basel) 12:N/A (2022). PubMed: 35745286
  • Hyldbakk A  et al. Identification of novel cyanoacrylate monomers for use in nanoparticle drug delivery systems prepared by miniemulsion polymerisation - A multistep screening approach. Int J Pharm X 4:100124 (2022). PubMed: 35898812
  • Kong X  et al. Transcranial Direct-Current Stimulation Regulates MCT1-PPA-PTEN-LONP1 Signaling to Confer Neuroprotection After Rat Cerebral Ischemia-Reperfusion Injury. Mol Neurobiol 59:7423-7438 (2022). PubMed: 36190692
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-9 of 9 Abreviews or Q&A

Question

what is the difference between both kits?
can one them to be used with tissue culture supernatant only (no cells)?

Read More

Abcam community

Verified customer

Asked on Nov 06 2012

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.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Abcam Scientific Support

Answered on Nov 06 2012

Question

I have discussed the situation with my PI and we would like a refund. We did a control experiment with the remainder of the kit and we found the same problem. How do we go about getting the refund?
Thank you,

Read More

Abcam community

Verified customer

Asked on Apr 10 2012

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.

Read More

Abcam Scientific Support

Answered on Apr 10 2012

Question

I spoke to you on the phone today (4/3/12) just before 2pm. We are having problems with an LDH assay and you and I were discussing how to analyze the data. I have attached a spreadsheet with the data and additional information. Please email or call me if you have any questions.
Thank you for your help,

Read More

Abcam community

Verified customer

Asked on Apr 06 2012

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.

Read More

Abcam Scientific Support

Answered on Apr 06 2012

Question

Dear technical team,

Our customer would like to test LDHassay with tissue sample.
https://www.abcam.com/LDH-Cytotoxicity-Assay-Kit-400-assays-ab65391-support.html

Is it possible with ab65391?

Let me know.


Best regards

Read More

Abcam community

Verified customer

Asked on Jan 27 2012

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.

Read More

Abcam Scientific Support

Answered on Jan 27 2012

Question

Does this LDH-Cytotoxicity Assay Kit suitable for all species?Including rat and mouse cells.

Read More

Abcam community

Verified customer

Asked on Dec 20 2011

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.    

Read More

Abcam Scientific Support

Answered on Dec 20 2011

Question

Dear technical team,   Our customer would like to purchase LDH-Cytotoxicity Assay Kit and I have searched two products on the website.   But I don't know well what the differrences between them, ab65391 and ab65393.   Let me know, please.   Thank you in advance.  

Read More

Abcam community

Verified customer

Asked on Dec 07 2011

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.

Read More

Abcam Scientific Support

Answered on Dec 07 2011

Question

Which of these 2 kits would be suitable for a use on HeLa cells as well as mucus?  

Read More

Abcam community

Verified customer

Asked on Nov 23 2011

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.      

Read More

Abcam Scientific Support

Answered on Nov 23 2011

Question

I'm pretty interested in buying your LDH-Cytotoxicity Assay Kit (400 assays) (ab65391). But before placing the order, I want to make sure if we can read the products. We have a plate reader, which is BioTek Synergy 2. Is it good for reading the samples?

Read More

Abcam community

Verified customer

Asked on Nov 22 2011

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.

Read More

Abcam Scientific Support

Answered on Nov 22 2011

Question

What is the difference between the LDH cytotoxicity assay kits ab65393 and ab65391?

Read More

Abcam community

Verified customer

Asked on Aug 04 2011

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.

Read More

Abcam Scientific Support

Answered on Aug 04 2011

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