Overview

  • Product name

    L-Lactate Assay Kit (Colorimetric/Fluorometric)
    See all L-Lactate kits
  • Detection method

    Colorimetric/Fluorometric
  • Sample type

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

    Quantitative
  • Sensitivity

    > 0.001 mM
  • Range

    0.001 mM - 10 mM
  • Assay time

    0h 40m
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    L-Lactate Assay Kit (Colorimetric/Fluorometric) ab65330 provides a convenient means for detecting L(+)-Lactate in biological samples such as blood, cells, culture mediums, fermentation mediums, etc. There is no need for pretreatment or purification of samples.


    In the lactate assay protocol, lactate specifically reacts with an enzyme mix to generate a product, which interacts with a lactate probe to produce color (570 nm) and fluorescence (at Ex/Em = 535/587 nm).


    Lactate assay protocol summary:
    - add samples (deproteinized) and standards to wells
    - add reaction mix and incubate for 30 min at room temp
    - analyze with microplate reader

  • Notes

    L(+)-Lactate is the major stereo-isomer of lactate formed in human intermediary metabolism and is present in blood. D(-)-Lactate is also present (see D-Lactate assay kits) but only at about 1-5% of the concentration of L(+)-Lactate.

    Alternative L-Lactate assay kits offer different readout modes/wavelengths and sensitivity/range:
    - this kit L-Lactate assay ab65330: colorimetric 570 nm, fluorometric Ex/Em 535/587 nm, range 0.001 mM - 10 mM
    - our most popular L-Lactate assay ab65331: colorimetric 450nm, range 0.02 mM - 10 mM
    L-Lactate assay ab169557: fluorometric Ex/Em 535/587 nm, range 0.2 µM - 50 µM. 

    Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests
    L(+)-Lactate Standard (100 nmol/µl) Yellow 1 x 100µl
    Lactate Assay Buffer WM 1 x 25ml
    Lactate Enzyme Mix (lyophilized) Green 1 vial
    Lactate Probe (in DMSO, anhydrous) Red 1 x 200µl
  • Research areas

  • Relevance

    Lactate (CH3CH(OH)COO-) plays important roles in many biological processes. Abnormal high concentration of lactate has been related to disease states such as diabetes and lactate acidosis, etc. L(+)-Lactate is the major stereoisomer of lactate formed in human intermediary metabolism and is present in blood. The lactate to pyruvate ratio reflects the redox state of the cell and describes the balance beween NAD+ and NADH, which is dependent on the interconversion of lactate and pyruvate via lactate dehydrogenase (LDH).

Images

  • HaCaT cells are grown in normoxia for 3 days after infection and medium was tested to determine lactate concentration using L-Lactate assay kit (ab65330). HaCaT cells were infected with AdGFP-18E2 or AdGFP, transfected with Ctrl or HIF-1α siRNA.

  • Relative signal (RFU) in unfiltered human plasma (dilution 1:200) and mouse urine (dilution 1:200), comparing L-lactate signals with background readings ((-); no enzyme) after 10 minutes of incubation (duplicates +/- SD).

  • Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Lactate measured in cell culture supernatants showing quantity (µmol) per mL of tested sample. Samples were diluted 40-160 fold and measured fluorometrically.

  • Lactate measured in biological fluids showing quantity (µmol) per mL of tested sample. Samples were diluted 10-40 fold and measured fluorometrically.

Protocols

References

This product has been referenced in:

  • Schnack L  et al. Mechanisms of Trained Innate Immunity in oxLDL Primed Human Coronary Smooth Muscle Cells. Front Immunol 10:13 (2019). Read more (PubMed: 30728822) »
  • Girardot T  et al. Intra-cellular lactate concentration in T lymphocytes from septic shock patients - a pilot study. Intensive Care Med Exp 6:5 (2018). Read more (PubMed: 29404815) »
See all 17 Publications for this product

Customer reviews and Q&As

1-10 of 21 Abreviews or Q&A

Question
Answer



I can confirm that we suggest to start with 10mg of tissue (muscle for example) or 10 million cells if using single cells.

This will vary widely depending on the tissue or cell type.

If you are using a cell type where the expected lactate amount is un-known, we recommend to also run a positive control cell along the side.

I hope this information is helpful and wish you good luck with your research.

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Answer



I can confirm that the assay buffer and the standard need to be stored at -20C as stated on the datasheet.

However, if assays will be run in small batches, we can recommend aliquoting these components and storing them to prevent freeze-thaw cycles.

Read More

Answer

Yes, our ab65330 L-Lactate Assay Kit (Fluorometric) can be used for colorimetric detection. The difference this kit has with the ab65331 L-Lactate Assay Kit (Colorimetric) is indeed mainly the probe, but the detection limits are also different for both. ab65330: 0.001-10 mM and ab65331 : 0.02 mM – 10 mM.

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Answer

Thank you for contacting us. It should have mentioned in the protocol the incubation time is 30 minutes. We will be sure to correct the protocol.


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

Answer

Thank you for contacting Abcam.
Since the lactate dehydrogenase (LDH) in tissue lysates degrades lactate, be sure to keep your samples at -80ºC. Below is our recommended homogenization protocol:
1. Homogenize the tissue samples (should contain at least 2 million cells) in the assay buffer that is provided for 10-50 passes on ice.
2. Centrifuge the sample and collect the supernatant.
3. Load the supernatant onto a 10kda spin column (ab93349 https://www.abcam.com/10kd-spin-column-ab93349.html) for deproteinization to remove LDH. Use the eluate for your subsequent assays.
To make sure that your readings from these samples are within the standard curve, we suggest using several doses of your sample when running your assay.
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 enquiry.

We would recommend to use the assay buffer provided with ab65330 L-Lactate Assay Kit (Fluorometric) for cells. This buffer contains some proprietary ingredients which are crucial for the working efficiency of this assay. Using just Triton-X-100 will compromise that efficiency and the samples will not work so well in the assay.

I hope this information is helpful to you. If you have any further questions, pleasedo not hesitate to contact us.

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Question
Answer

Thank you for contacting Abcam earlier today.

To prepare an intracellular lysate to use in the assay, we recommend using at least 2 x 10^6 cells. Then homogenize the cells in the assay buffer that is provided and centrifuge it gently to get rid of the foam/air bubbles. To make sure that your readings from these samples are within the standard curve, we suggest using several doses of your sample when running your assay.

If there is anything else that I can help you with, please let me know.

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Answer

Gracias por contactarnos.

El kit de detección de Lactato debe mantenerse a -20C. Almacenado a esta temperatura y evitando ciclos de congelación y descongelación, el kit dura hasta la fecha indicada en la caja del kit, que suelen ser un par de años.

Es importante sin embargo que se tenga en cuenta el periodo de validez de los distintos componentes del kit una vez se han descongelado y disuelto.

Espero que esta información sea de ayuda. En caso contrario, por favor, no dudes en volvernos a contactar.

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Answer

Thank you for contacting us.

I am not familiar with the contents of MRSB media, but in general cell culture supernatants will be compatible with these kits. If the media contains serum, we recommend deproteinizing the samples using our 10 kDa cutoff spin columns (ab93349) to prevent degradation of the lactate. Phenol red is generally not a problem because the samples will be substantially diluted, but you can always add some diluted media to your blank wells to control for any additional background.

I hope this helps, please let me know if you need any additional information or assistance.

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1-10 of 21 Abreviews or Q&A

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