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    products/assay-kits/l-lactate-assay-kit-colorimetricfluorometric-ab65330.pdf

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L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (2)Q&A (21)References (229)

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Functional studies - ab65330
  • Functional Assay: L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)
  • Functional Studies - L-Lactate Assay Kit (ab65330)
  • Functional Studies - L-Lactate Assay Kit (ab65330)
  • Functional Studies - L-Lactate Assay Kit (ab65330)
  • Functional Studies - L-Lactate Assay Kit (ab65330)

Key features and details

  • Assay type: Quantitative
  • Detection method: Colorimetric/Fluorometric
  • Platform: Microplate reader
  • Assay time: 40 min
  • Sample type: Cell culture supernatant, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Lysate, Urine
  • Sensitivity: 0.001 mM

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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

    This product is manufactured by BioVision, an Abcam company and was previously called K607 Lactate Colorimetric/Fluorometric Assay Kit. K607-100 is the same size as the 100 test size of ab65330.

    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 2000 tests
    L(+)-Lactate Standard (100 nmol/µl) Yellow 1 x 100µl 20 x 100µl
    Lactate Assay Buffer WM 1 x 25ml 20 x 25ml
    Lactate Enzyme Mix (lyophilized) Green 1 vial 20 vials
    Lactate Probe (in DMSO, anhydrous) Red 1 x 200µl 20 x 200µl
  • Research areas

    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Sugar Assays
  • 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

  • Functional studies - ab65330
    Functional studies - ab65330Image from Lai D., PLiS One 8(9), Fig 6C. Doi: 10.1371/journal.pone.0075625. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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.

  • Functional Assay: L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)
    Functional Assay: L-Lactate Assay Kit (Colorimetric/Fluorometric) (ab65330)

    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).

  • Functional Studies - L-Lactate Assay Kit (ab65330)
    Functional Studies - L-Lactate Assay Kit (ab65330)

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

  • Functional Studies - L-Lactate Assay Kit (ab65330)
    Functional Studies - L-Lactate Assay Kit (ab65330)

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

  • Functional Studies - L-Lactate Assay Kit (ab65330)
    Functional Studies - L-Lactate Assay Kit (ab65330)

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

  • Functional Studies - L-Lactate Assay Kit (ab65330)
    Functional Studies - L-Lactate Assay Kit (ab65330)

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

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (229)

Publishing research using ab65330? Please let us know so that we can cite the reference in this datasheet.

ab65330 has been referenced in 229 publications.

  • Davern M  et al. Acidosis significantly alters immune checkpoint expression profiles of T cells from oesophageal adenocarcinoma patients. Cancer Immunol Immunother 72:55-71 (2023). PubMed: 35708739
  • Wang CS  et al. NBR2/miR-561-5p/DLC1 axis inhibited the development of multiple myeloma by activating the AMPK/mTOR pathway to repress glycolysis. Neoplasma 69:1165-1174 (2022). PubMed: 35951458
  • Ding D  et al. Non-canonical function of an Hif-1a splice variant contributes to the sustained flight of locusts. Elife 11:N/A (2022). PubMed: 36039636
  • Shyam R  et al. Rescue of the Congenital Hereditary Endothelial Dystrophy Mouse Model by Adeno-Associated Viruse-Mediated Slc4a11 Replacement. Ophthalmol Sci 2:N/A (2022). PubMed: 36051248
  • Chen J  et al. Activated Hepatic Stellate Cells Promote the M1 to M2 Macrophage Transformation and Liver Fibrosis by Elevating the Histone Acetylation Level. Dis Markers 2022:9883831 (2022). PubMed: 36133436
View all Publications for this product

Customer reviews and Q&As

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1-10 of 23 Abreviews or Q&A

Lactate assay is quick and sensitive

Excellent Good 4/5 (Ease of Use)
Abreviews
Abreviews
abreview image
We used this lactate assay to detect lactate in supernatants and tissue samples and the kit was very simple to use and very fast. Samples likely need to be diluted if deproteinized, and if the concentration range is unknown, an additional one or two standards above or below the recommended standards can be used to cover a greater dynamic range.
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 Jan 16 2020

L-lactate assay kit ab65330

Good Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
I used the lactate assay kit ab65330 for colorimetric measurement of lactate in cell supernatant. After collection, samples are deproteinated by filtering through 10kDa filter. Samples are then diluted 1/25 and analysed according to protocol. The kit is very easy to use, fast, and also works on frozen supernatant (store samples at -80C after deproteination).
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.

Mandy Grootaert

Verified customer

Submitted Jan 06 2020

Question

sample: cell lysate, how many cells as minimum?

Read More

Abcam community

Verified customer

Asked on May 29 2013

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.

Read More

Abcam Scientific Support

Answered on May 29 2013

Question

I am using the L-Lactate Assay Kit (Fluorometric) which has to be stored at -20*C. I was wondering if it is possible to store the buffer separately at a different temperature like 4*C as it takes longer to defrost?


Read More

Abcam community

Verified customer

Asked on Sep 13 2013

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

Janina Burkhardt

Abcam Scientific Support

Answered on Sep 13 2013

Question

Customer is wondering if this kit can also be used for colorometric detection. On our datasheet we have it listed as being able to produce color at 570 nm as well as fluorescence. With your other L-Lactate Assay Kit (colorometric), what is the difference? Is it the probe?

Read More

Abcam community

Verified customer

Asked on Jun 11 2013

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.

Read More

Abcam Scientific Support

Answered on Jun 11 2013

Question

I’ve been using your L-Lactate assay kit (ab65330), Glucose assay kit (ab65333) and Free Glycerol detection kit (ab65337) to collect data. I had no problem with measurement except 1 series of samples. I would like to modify the condition to see whether it gets better or not. I would appreciate, if you could help me to figure it out.



In the assay, we mix the probe and enzyme mix into assay buffer. I already tried to dilute “Probe” concentration. I would like to try to change the portion of “Probe” and “Enzyme Mix”. Could you please tell me what is in the “Probe” and “Enzyme Mix” in each of 3 kits? Does “probe” contain both fluorescent dye and NAD (or other materials)? Does Enzyme Mix contain only Lactate dehydrogenase or other materials too? It would be a great help to modify the portion of each materials (Dilution and supplementation).

Read More

Abcam community

Verified customer

Asked on Mar 05 2013

Answer

I cannot give the exact compositions of the enzyme mixes in these kits, but the probes are just pure probes in a solvent. They do not have any other components. The enzyme mixes are obviously a combination of multiple enzymes. It does not have any florescent dyes or co-factors.

Read More

Abcam Scientific Support

Answered on Mar 05 2013

Question

If so, how long should we have incubated the samples before reading the plate? Should we rerun our experiment in order to obtain data that we can accurately

Read More

Abcam community

Verified customer

Asked on Dec 19 2012

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

Abcam Scientific Support

Answered on Dec 19 2012

Question

Product code: 65330
Lot number: XXXXXX
º
Inquiry: Hi, We just bought the L-Lactate assay kit and I cannot find in the assay protocol how should I homogenize my tissue samples (hypothalamus). I will apreciate information about this issue in order to process the samples. Thanks

Read More

Abcam community

Verified customer

Asked on Nov 26 2012

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.
Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Abcam Scientific Support

Answered on Nov 27 2012

Question

Product code: 65330

Inquiry: I would like to try the lactate assay kit ≠ab65330 for determination of intracellular lactate content. I have to work in 96well plate and I want to know whether using a standard lysis buffer (say with 0.5% TritonX-100) would affect the assay readout. Thanks

Read More

Abcam community

Verified customer

Asked on Sep 26 2012

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.

Read More

Abcam Scientific Support

Answered on Sep 26 2012

Question

How to prepare cell culture extracts for use in this assay?

Read More

Abcam community

Verified customer

Asked on Sep 18 2012

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.

Read More

Abcam Scientific Support

Answered on Sep 18 2012

1-10 of 23 Abreviews or Q&A

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