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    products/assay-kits/l-lactate-assay-kit-colorimetric-ab65331.pdf

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L-Lactate Assay Kit (Colorimetric) (ab65331)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (2)Q&A (60)References (259)

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L-Lactate Assay Kit (Colorimetric) ab65331
  • ab65331 L-Lactate Assay Kit (Colorimetric)
  • L-Lactate Assay Kit (Colorimetric) (ab65331)
  • ab65331 L-Lactate Assay Kit (Colorimetric)
  • L-Lactate Assay Kit (Colorimetric) (ab65331)

Key features and details

  • Assay type: Quantitative
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Assay time: 30 min
  • Sample type: Cell culture supernatant, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Lysate, Urine
  • Sensitivity: 0.02 mM

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Overview

  • Product name

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

    Colorimetric
  • Sample type

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

    Quantitative
  • Sensitivity

    0.02 mM
  • Range

    0.02 mM - 10 mM
  • Assay time

    0h 30m
  • Product overview

    L-Lactate Assay Kit (Colorimetric) (ab65331) uses an assay protocol where lactate is oxidized by lactate dehydrogenase to generate a product which interacts with a probe to produce a color (λmax = 450 nm).


    The kit detects L(+)-Lactate in biological samples such as serum or plasma, cells, tissues, cell culture and fermentation media. 


    Lactate assay protocol summary:
    - add samples 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 K627 Lactate Colorimetric Assay Kit II. K627-100 is the same size as the 100 test size of ab65331.

    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.

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

    How other researchers have used L-Lactate Assay Kit ab65331

    This Lactate assay kit has been used in publications in a variety of sample types, including:
    - Human: THP-1 cell lysates1, MDA-MB-231 and HepG2 cell culture lysates2, cell culture supernatant (HepG2, A549, Huh7, PC3, LN229, HeLa)3, brain tissue4
    - Mouse: brown adipose tissue lysate5, thymic lymphoma tissue6, cell culture supernatant7, T cell primary cell culture supernatants8, serum9, serum and muscle10
    - Bovine: cumulus cell culture supernatant11 

    References: 1 - Tran UT and Kitami T 2019; 2 - Cui J et al 2019; 3 - Rodriguez ML et al 2018, Chen Y et al 2018, Zhang D et al 2018, Caino MC et al 2017, Birkenmeier K et al 2015; 4 - Sullivan RC et al 2019; 5 - Jeong JH et al 2018; 6 - Vara-Ciruelos D et al 2019; 7 - Fiorenzano et al 2016; 8 - Menk AV et al 2018; 9 - Deng W et al 2019, Guglielmetti C et al 2017, Kang R et al 2016; 10 - Kim HY et al 2016; 11 - Sinha et al 2017

  • 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 Substrate Mix Red 1 vial
  • 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).

Associated products

  • Related Products

    • 10kD Spin Column (ab93349)

Images

  • L-Lactate Assay Kit (Colorimetric) ab65331
    L-Lactate Assay Kit (Colorimetric) ab65331Image from Kelly CE et al., PLoS Biol 11(4), fig2d. oi: 10.1371/journal.pbio.1001538. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Plasma lactate concentrations were determined using L-Lactate assay kit (ab65331) in Ark2C+/+ and Ark2C−/− (Arkadia-like gene) mice.

  • ab65331 L-Lactate Assay Kit (Colorimetric)
    ab65331 L-Lactate Assay Kit (Colorimetric)

    Linearity of dilution: concentration of L-Lactate in differently diluted (X-axis) biological samples, demonstrating a linearity of 89%-111% (concentrations corrected for by factor of dilution; duplicates; +/- SD).

  • L-Lactate Assay Kit (Colorimetric) (ab65331)
    L-Lactate Assay Kit (Colorimetric) (ab65331)

    Relative signal (RFU) in unfiltered human plasma (dilution 1:8), comparing L-lactate signals with background reading (no enzyme) after 10 minutes of incubation (duplicates +/- SD).

  • ab65331 L-Lactate Assay Kit (Colorimetric)
    ab65331 L-Lactate Assay Kit (Colorimetric)

    Standard curve with background signal subtracted (duplicates; +/- SD).

  • L-Lactate Assay Kit (Colorimetric) (ab65331)
    L-Lactate Assay Kit (Colorimetric) (ab65331)
    Lactate Standard Curve. The assay is performed following the kit (ab65331) protocol.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (259)

Publishing research using ab65331? Please let us know so that we can cite the reference in this datasheet.

ab65331 has been referenced in 259 publications.

  • Das A  et al. Systemic ablation of vitamin D receptor leads to skeletal muscle glycogen storage disorder in mice. J Cachexia Sarcopenia Muscle 13:467-480 (2022). PubMed: 34877816
  • Vultaggio-Poma V  et al. Extracellular ATP is increased by release of ATP-loaded microparticles triggered by nutrient deprivation. Theranostics 12:859-874 (2022). PubMed: 34976217
  • Gao L  et al. Trophoblast-derived Lactic Acid Orchestrates Decidual Macrophage Differentiation via SRC/LDHA Signaling in Early Pregnancy. Int J Biol Sci 18:599-616 (2022). PubMed: 35002512
  • Dai B  et al. Macrophages in epididymal adipose tissue secrete osteopontin to regulate bone homeostasis. Nat Commun 13:427 (2022). PubMed: 35058428
  • Harrison MAA  et al. Metabolic Reprogramming of Glioblastoma Cells during HCMV Infection Induces Secretome-Mediated Paracrine Effects in the Microenvironment. Viruses 14:N/A (2022). PubMed: 35062307
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-10 of 62 Abreviews or Q&A

Lactate assay is very quick and easy to use

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
The lactate assay is very simple and easy to use. I used it to measure levels of lactate in cell supernatant and was able to use fresh supernatant without any cleaning/deproteinizing. The standard curve and sensitivity are also good. Overall, the kit is great.
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.

Robert Yuan

Verified customer

Submitted Aug 28 2019

L-lactate assay kit

Good Good 4/5 (Ease of Use)
Abreviews
Abreviews
abreview image
The assay kit is a fast and easy way to quantify lactate concentrations. It works well for whole cell lysates (WCE). HCT116 cells were cultivated for 48 and 72 hrs in DMEM and sonified in assay buffer. The lactate concentration is displayed in the enclosed figure, dependent on the amount of WCE.
The kit only got 4 stars because it does not work in DMEM thus the supernatant of the cells could not be analysed. 10 µl DMEM in combination with assay buffer and the reaction mix gave absorbance values over 1.
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.

Mr. Christian Marx

Verified customer

Submitted Jul 23 2013

Question

Bonjour,



Merci pour les renseignements,



Concernant les dosages nous avons réessayé une fois mais sans amélioration des résultats. Par la suite, nous avons fait doser les échantillons dans un service de Biochimie qui utilisent des kits d'un autre fabricant et cette fois-ci les résultats étaient corrects.



Merci beaucoup,



Je reste à votre disposition pour toute information complémentaire

Read More

Abcam community

Verified customer

Asked on Jul 04 2012

Answer

Merci pour cette information. Je suis très désolée que ces kits n'ont pas permis de faire les dosages attendus. Je vais volontiers organiser une note de crédit ou un remboursement pour vous. Alternativement je peux aussi organiser l'envoie de deux kits de remplacement, mais je pense que vous préférez probablement la note de crédit.Est-ce que vous pouvez me confirmer quel 'est la solution que vous préférez?

Merci beaucoup. Je reste à votre disposition pour toute information complémentaire.

Read More

Abcam Scientific Support

Answered on Jul 04 2012

Question

I see that assay buffer can be used to extract cells for use with this kit. But there is not protocol describing how to use the assay buffer to produce the cell extract. Please provide a protocol.

Read More

Abcam community

Verified customer

Asked on Apr 09 2014

Answer

For cell samples: Start with ˜2x106 cells, suspend the cell pellet 500 μl (or ˜4 volumes) of the Assay Buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. Spin down the sample and collect the supernatant. For enzymztic assay kits, it may be useful to deproteinate samples now by loading the supernatant into a 10kda spin column for deproteinzation. (Use the eluate for your subsequent assays.) Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve.

Read More

Jeremy Kasanov

Abcam Scientific Support

Answered on Apr 09 2014

Question

I was wondering if Phenol Red would interfere with the Lactate assay kit? Although, can I make the standard curve up in the correspondent complete cell culture media (either RPMI/10%FCS+Phenol Red or DMEM/10%FCS+Phenol Red) or does it has to be in the Lactate assay buffer?

Read More

Abcam community

Verified customer

Asked on Feb 12 2014

Answer



I can confirm that this kit (ab6533:L-Lactate Assay Kit) uses 450 nm absorbance reading and hence the color should not affect the measurements.

For those kits that measure absorbance at 570 nm, the red color could potentially interfere but only if a lot of media is used. Typically up to 50 ul medium per well does not affect the absorbance reading since the solution gets diluted with reaction mix.


Read More

Tanya Bagrij

Abcam Scientific Support

Answered on Feb 12 2014

Question

I have a question about this kit. Can't one just measure the increase in NADH produced at by measuring increased absorbance at OD340? Doesn't this correspond directly to lactate levels? Why is it necessary to measure production of formazon at OD450 intead?

Read More

Abcam community

Verified customer

Asked on Dec 10 2013

Answer

Yes, theoretically it can be done by monitoring the changes in NADH absorption at 340 nm.

However, measuring at this OD requires expensive quartz microplates/cuvettes.
Enzymatic assays like ab65331 are a better way to get more accurate measurements because the signal is more specific and magnified, allowing better sensitivity and accuracy.

Read More

Jeremy Kasanov

Abcam Scientific Support

Answered on Dec 10 2013

Question

What is the standard protein host species?

Read More

Abcam community

Verified customer

Asked on Nov 15 2013

Answer

The standard is 98% pure sodium-L-lactate (chemical, not animal derived).

Read More

達偉 鄭

Abcam Scientific Support

Answered on Nov 15 2013

Question

Inquiry: Hi, In your other kits (ab65336) and (ab65620) we have used, you list a volume to use for tissue extractions. In the L-lactate kit you do not and it would be very helpfull to know the tissue sample size and recommended volume. The tissue in question is rat muscle and liver. Thank you

Read More

Abcam community

Verified customer

Asked on Sep 16 2013

Answer

Thank you for your enquiry.

For this kit (ab65331) 10-100mg tissue can be used depending upon the lactate in the tissue. This will require some optimization depending on the sample. We recommend using different dilutions of the homogenate so that the best dilution for results within the linear range of the std curve can be found.

I. hope this information is useful to you. Please do not hesitate to contact us if you have any further questions.

Read More

Sam Washer

Abcam Scientific Support

Answered on Sep 16 2013

Question

Hi,
we've been trying to assay extracellular lactate (in the medium) conditioned by Drosophila cells. We use Schneider's medium + 10% FBS. We've used your ab65331 kit and measured the medium with no cells, and then medium conditioned for 24hrs from cells. The medium is put over 10kDa spin columns (Ab93349) to rid it of HMW factors. However, the results indicate that there is quite a bit of lactate in the medium in the absence of cells (the wells were treated the same way as the medium that was taken from the cell cultures). I cannot find any mention of lactate or pyruvate in the Schneider medium formulation. Can you help me understand where the lactate is coming from?

Read More

Abcam community

Verified customer

Asked on Jul 01 2013

Answer

Thank you for contacting us.

Fetal Bovin Serum contains LDH and will create high background levels. Using a low percent FBS (0.25-1%) or serum free media will reduce this background.

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

Heather Allen

Abcam Scientific Support

Answered on Jul 01 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

1-10 of 62 Abreviews or Q&A

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