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    products/assay-kits/ldh-assay-kit--lactate-dehydrogenase-assay-kit-colorimetric-ab102526.pdf

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LDH Assay Kit / Lactate Dehydrogenase Assay Kit (Colorimetric) (ab102526)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (1)Q&A (19)References (195)

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LDH assay on mouse skeletal muscle
  • Functional Studies - Lactate Dehydrogenase Colorimetric Assay Kit (ab102526)
  • Functional Studies - Lactate Dehydrogenase Colorimetric Assay Kit (ab102526)
  • Functional Studies - Lactate Dehydrogenase Colorimetric Assay Kit (ab102526)

Key features and details

  • Assay type: Enzyme activity (quantitative)
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 30 min
  • Sample type: Cell culture supernatant, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine
  • Sensitivity: 1 mU/ml

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Overview

  • Product name

    LDH Assay Kit / Lactate Dehydrogenase Assay Kit (Colorimetric)
    See all Lactate Dehydrogenase kits
  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts
  • Assay type

    Enzyme activity (quantitative)
  • Sensitivity

    > 1 mU/ml
  • Range

    1 mU/ml - 100 mU/ml
  • Assay time

    0h 30m
  • Product overview

    Lactate Dehydrogenase (LDH) Assay Kit (Colorimetric) ab102526 quantifies LDH activity in a variety of samples such as serum or plasma, tissue, cells, and culture medium.


    In the LDH assay protocol, LDH reduces NAD to NADH, which then interacts with a specific probe to produce a color (OD max = 450 nm). The kit can detect 1 - 100 mU/mL of LDH directly in samples. The assay is quick, convenient, and sensitive.


    LDH assay protocol summary:
    - add samples and standards to wells
    - add reaction mix
    - analyze every 2-3 min for at least 30 min with microplate reader in kinetic mode at 37ºC

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called K726 Lactate Dehydrogenase Activity Colorimetric Assay Kit. K726-500 is the same size as the 500 test size of ab102526.

    To measure LDH release into cell culture medium from cultured cells in a cytoxicity experiment, we recommend LDH assay kit ab65393, which is designed specifically as a cytotoxicity assay.

    This LDH assay kit ab102526 is designed in a more flexible format for use with a variety of sample types (this includes cell culture medium, and this kit may be used in cytotoxicity assays). This flexible format LDH assay kit is also available as a fluorometric LDH assay kit ab197000.

     

    How other researchers have used LDH Assay Kit ab102526

    This LDH assay kit has been used in publications in a variety of sample types, including:
    - Human: serum1
    - Mouse: kidney tissue2, kidney epithelial cell line lysates3, muscle tissue4, serum2,5, vaginal lavage fluid6, liver tissue7
    - Rat: H8C2 cell lysates8

    It has also been used in LDH release cytotoxicity cell culture assays including in human hippocampal neuronal cultures9, THP-1 cells10, primary chondrocyte cells on a membrane scaffold11, primary lymphocytes12, ovarian cancer cells co-cultured with mouse splenocytes in a cytotoxic T lymphocyte assay13

    References: 1-Zhang Z et al 2018, Solivio MJ et al 2016; 2-Niles DJ et al 2018; 3-Tanner LB et al 2018; 4-Tam BT et al 2018; 5-Kim HY et al 2016; 6-Nash EE et al 2016; 7-Lai C et al 2018; 8-Zhou Z et al 2018; 9-Wang Q et al 2019; 10-Alimov I et al 2019; 11-Stocco TD et al 2019; 12-Dreisig K et al 2018; 13-Sun Y et al 2018

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 500 tests
    LDH Assay Buffer NM 1 x 50ml
    LDH Positive Control  Red 1 vial
    NADH Standard I Yellow 1 vial
    Substrate Mix Amber 1 vial
  • 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
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Lipid Metabolism Kits
    • Insulin signaling
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Lipid Metabolism Kits
    • Lipid metabolism
    • 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

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

Images

  • LDH assay on mouse skeletal muscle
    LDH assay on mouse skeletal muscleTam BT et al. 2018 Sci Rep 8:3689 Use licensed under: http://creativecommons.org/licenses/by/4.0/

    Tam BT et al. used the LDH assay kit on mouse skeletal muscle tissue lysates comparing pressure-induced compressed and control samples across a range of knockout mice lines.

  • Functional Studies - Lactate Dehydrogenase Colorimetric Assay Kit (ab102526)
    Functional Studies - Lactate Dehydrogenase Colorimetric Assay Kit (ab102526)

    Kinetic profiles of approx 0.5 mU of a sample of pure LDH (Positive Control) and 2 μL frozen human serum using buffer as a background control.

  • Functional Studies - Lactate Dehydrogenase Colorimetric Assay Kit (ab102526)
    Functional Studies - Lactate Dehydrogenase Colorimetric Assay Kit (ab102526)

    NADH Standard Curve.

  • Functional Studies - Lactate Dehydrogenase Colorimetric Assay Kit (ab102526)
    Functional Studies - Lactate Dehydrogenase Colorimetric Assay Kit (ab102526)Image from Xu Y et al., PLoS One. 2014;9(6):e100127. Fig 2(B).; doi: 10.1371/journal.pone.0100127. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Cellular necrosis was measured as LDH release after cisplatin treatment for 24 h and 48 h using ab102526. The release of LDH was apparently increased at 24 and 48 h after cisplatin treatment.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (195)

Publishing research using ab102526? Please let us know so that we can cite the reference in this datasheet.

ab102526 has been referenced in 195 publications.

  • Chen X  et al. Identification of HPCAL1 as a specific autophagy receptor involved in ferroptosis. Autophagy 19:54-74 (2023). PubMed: 35403545
  • Valdiviezo A  et al. Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models. Toxics 10:N/A (2022). PubMed: 36287846
  • Li D  et al. The effect of a spontaneous induction prophage, phi458, on biofilm formation and virulence in avian pathogenic Escherichia coli. Front Microbiol 13:1049341 (2022). PubMed: 36452923
  • Ramos-Martinez E  et al. Enhanced Activity of NLRP3 Inflammasome in the Lung of Patients with Anti-Synthetase Syndrome. Cells 12:N/A (2022). PubMed: 36611853
  • Sharifiaghdam Z  et al. Curcumin-coated gold nanoparticles attenuate doxorubicin-induced cardiotoxicity via regulating apoptosis in a mouse model. Clin Exp Pharmacol Physiol 49:70-83 (2022). PubMed: 34449914
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-10 of 20 Abreviews or Q&A

Abreview forLDH Assay Kit ( ab102526)

Good Good 4/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Set up 48-well plate using MM1.S cells.
Collect 60 ul of supernatant from each well and treat with different concentrations of PEG (600-3000)
At final timepoint, collect all cells, spin celle to pellet and add 100 ul of LDH lysis buffer.
Reaction:
50 ul of supernatant+50 ul of LDH substrate—RT,30 min
40 ul of PBS+10 ul of cell lysate+50 ul of LDH substrate—RT,30 min
Add 50 ul of stop solution.
Measuren absorbance at 490 nm
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 02 2018

Question

I sthe kit ab65393 suitable for tissue culture supernatant.

Read More

Abcam community

Verified customer

Asked on Mar 11 2014

Answer

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 we cannot confirm its specificity. More likely it will work with tissue extracts as well. Please note you might need to optimize the amount of tissue by trying several dilutions.

Read More

Padamjeet Singh

Abcam Scientific Support

Answered on Mar 11 2014

Question

I have kit for LDH # 102526 and I am trying to use it to measure the LDH on lung tissue.

I used 50mg of tissue and homogenized in 250 μl buffer, then I spun down and used the supernatant. For my assay I diluted 1:10 the original supernatant and then I used 5μl for the assay.



Read More

Abcam community

Verified customer

Asked on Mar 25 2013

Answer

Hello!
Based on the info you sent you basically diluted 1:10 once witht he original supernatant and then 1:10 again by using 5 ul plus assay buffer to make 50 ul total, so your final dilution is 1:100. Please let me know if you have any further questions.

Read More

Abcam Scientific Support

Answered on Mar 25 2013

Question

LDH = (B*Sample Dilution)/((T2-T1)*V) = mU/ml

My question is, if we use 5ul of sample diluted to 50ul then what values should we use for the sample dilution and volume? The grad student running the assay used 10 for the sample dilution and 0.005 (5ul) as the sample volume, but I feel like this might be accounting for the dilution twice.

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Abcam community

Verified customer

Asked on Mar 07 2013

Answer


The dilution is the initial sample dilution. If you have 10 µl of a sample and are diluting it 100 times and then taking in 5 µl from that diluted sample into the well, your dilution factor would be 100. In this case dilution would be 1 since you are taking pure undiluted 5 µl of the sample into the wells, and the volume will be 0.005 ml.

Read More

Abcam Scientific Support

Answered on Mar 07 2013

Question

vorrei sapere se avete un kit per misurare la lattato deidrogenasi direttamente dal terreno delle cellule. Ho questo problema tecnico: le cellule sono fatte crescere su scaffold e non vorrei staccarle per ri-piastrarle in una multiwell, ma vorrei solamente prelevare il terreno e lì misurarci la lattato deidrogenasi rilasciata.

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Abcam community

Verified customer

Asked on Jan 18 2013

Answer


What I understand from your question is, that you would like a kit which measure lactate dehydrogenase produced from cells found in soil? In addition you would like to not detach the cells from the scaffold on which you are growing them to then replate them in the multi-well plate ready for the assay?
We have a kit ab102526 that measures Lactate Dehydrogenase activity by colorimetric means. This kit is suitable to use with cell culture supernatents. This means that you would have to grow your cells on separate dishes e.g on your scaffolds and then detach them to centrifuge at 10,000 x g for 15 min at 4°C. This way the supernatent can be collected and used for the assay.

Read More

Abcam Scientific Support

Answered on Jan 18 2013

Question

two vials (LDH substrate mix and NADH standard) were empty

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Abcam community

Verified customer

Asked on Jan 17 2013

Answer

Thank you for contacting us.

The two vials are actually lysophilized products. It does not say this on the datasheet, however the protocol booklet itself (which should have come with the kit and can also be downloaded from our website) gives you the instructions on its use. The substrate mix can be dissolved in 1ml ddH2O. The standard should be dissolved in 0.4 ml ddH2O to generate the 1.25 mM solution.


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 Jan 17 2013

Question

Buenas tardes,
Nos gustaría recibir el certificado analítico según lote, del siguiente producto:- Lactate Dehydrogenase (LDH) Assay Kit (Colorimetric); Ref: ab102526 Lot. XXXXX
Muchas gracias y un saludo,

Read More

Abcam community

Verified customer

Asked on Nov 13 2012

Answer

Gracias por contactarnos.

Adjunto a este mail envío el certificado de conformidad del kit ab102526.

Espero que os sea de utilidad, y en cualquier caso no dudéis en contactarnos para cualquier duda o sugerencia.

Read More

Abcam Scientific Support

Answered on Nov 13 2012

Question

Buenas tardes,
El pasado 28 de Septiembre recibí la oferta que adjunto. Se procedió a realizar el pedido de dicho producto y recibimos correctamente el kit.
Hoy hemos intentado probar dicho kit pero hemos tenido un problema. Los productos que venían en el kit estaban mal etiquetados y en estos momentos estamos dudando de si el reactivo que lleva cada producto es el correcto. Adjunto una fotografía tomada de dichos reactivos. El número de lote del kit es: XXXX. Nos gustaría que nos pudiesen dar una solución, ya sea mandando un comercial para poder comprobar que dichos reactivos están mal etiquetados, llamándome por teléfono o mandando otro kit.
Si necesitan alguna información adicional por favor pónganse en contacto conmigo.Muchas gracias.

Read More

Abcam community

Verified customer

Asked on Oct 22 2012

Answer

Te pido disculpas por la espera.

Siento mucho los inconvenientes causados por la confusión al etiquetar los componentes del kit. Estoy chequeando con mis compañeros responsables los lotes que tenemos actualmente en stock para confirmar que se han etiquetado como corresponde.

Una vez me lo verifiquen os mandaré un nuevo kit totalmente libre de cargos.

Muchas gracias por tu colaboración y paciencia con este asunto.

Read More

Abcam Scientific Support

Answered on Oct 22 2012

Question

Inquiry: your protocol for this assay contain wrong information. data analysis: equation is wrong! LDH activioty=Bxsample dilution/ (T2XT1)xV should be LDH activioty=Bxsample dilution/ (T2-T1)xV . T1 usually is 0! try getting a data from multiplying your data with 0!

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Abcam community

Verified customer

Asked on Oct 04 2012

Answer

Thank you for contacting us.

I am sorry that the equation provided in the protocol booklet of the Lactate Dehydrogenase (LDH) Assay Kit (Colorimetric), ab102526, does contain a typing error. the equation should read T2-T1 as you have pointed out. Thank you very much for bringing this to our attention. This will be corrected as soon as possible.

I am sorry for any inconvenience caused. If I can be of any further assistance, please do not hesitate to let me know.

Read More

Abcam Scientific Support

Answered on Oct 04 2012

Question

Hello: We are interested in using the colorimetric kit below for assays of culture media. Can you let us know if this kit requires that we use any special media specifically media with phenol red?
Lactate Dehydrogenase (LDH) Assay Kit (Colorimetric) (ab102526)
All the best

Read More

Abcam community

Verified customer

Asked on Aug 20 2012

Answer

Thanks for your enquiry regarding ab102526.

We have no specific recommendations for what kind of media you could use with this assay. I just looked it up online and it looks like phenol containing medium having ˜10% FBS will enhance the signals. Here is the abstract showing this:

Optimization, application, and interpretation of lactate dehydrogenase measurements in microwell determination of cell number and toxicity.

Wolterbeek HT, van der Meer AJ.

Source

Department of Radiation, Radionuclides, and Reactors, Faculty of Applied Sciences, University of Technology Delft, Delft, The Netherlands. H.T.Wolterbeek@tnw.tudelft.nl

Abstract

The lactate dehydrogenase (LDH) assay was addressed for its sensitivity, disturbances by foaming, and cell number and size. Cells were from a U-251 MG grade IV human glioblastoma brain tumor cell line used in 100-microl well volumes. Cells were counted by microscopy and Coulter counting; assays were LDH or trypan blue. The results indicate increased 490 nm signals (level, variance) by using phenol red or by increasing fetal bovine serum from 5% to 10%. The data also indicate that defoaming results in reduced variances ranging from a factor of 2 at 1-3 units of absorption, up to a factor of 4-5 at <1 units of absorption. Coulter counting indicated a decrease in cell volume with increasing end-point cell density, attributed to general shrinking at increasing density. In comparisons, total LDH was considered relative to both cell total volume and cell numbers. The result suggests that total LDH should be regarded as reflecting cell total volume rather than cell numbers. In a comparative Cu exposure test, signals of both LDH and a sodium salt of 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate (WST-1) decreased with increasing Cu supply, while bromodeoxyuridine signals remained largely unaffected. The data show the differences in responses in cell viability and proliferation, but, above all, indicate that LDH should be expressed on a per cell volume basis rather than per cell, to avoid the problem that mere density effects contribute to signals on compound or metal toxicity.

We are not certain if it would be true for this kit as well and that is why we don’t recommend any specific media.

Hope this information has been helpful for you.

Read More

Abcam Scientific Support

Answered on Aug 20 2012

1-10 of 20 Abreviews or Q&A

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