製品の概要

  • 製品名
    L-Lactate Assay Kit (Colorimetric)
    L-Lactate キット 製品一覧
  • 検出方法
    Colorimetric
  • サンプルの種類
    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Cell Lysate, Tissue Lysate
  • アッセイタイプ
    Quantitative
  • 検出感度
    0.02 mM
  • 検出範囲
    0.02 mM - 10 mM
  • 全工程の試験時間
    0h 30m
  • 製品の概要

    L-Lactate Assay Kit (Colorimetric) (ab65331) is an assay 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, culture and fermentation media. There is no need for pretreatment or purification of samples.


    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

  • 特記事項

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

    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.

  • 試験プラットフォーム
    Microplate reader

製品の特性

  • 保存方法
    Store at -20°C. Please refer to protocols.
  • 内容 ラベル 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
  • 研究分野
  • 関連性
    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).

関連製品

画像

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

  • 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).

  • 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).

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

  • Lactate Standard Curve. The assay is performed following the kit (ab65331) protocol.

プロトコール

参考文献

This product has been referenced in:
  • Jeong JH  et al. Intracellular glycolysis in brown adipose tissue is essential for optogenetically induced nonshivering thermogenesis in mice. Sci Rep 8:6672 (2018). Read more (PubMed: 29704006) »
  • Menk AV  et al. Early TCR Signaling Induces Rapid Aerobic Glycolysis Enabling Distinct Acute T Cell Effector Functions. Cell Rep 22:1509-1521 (2018). Read more (PubMed: 29425506) »
See all 36 Publications for this product

レビューと Q&A

1-10 of 62 Abreviews or Q&A

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

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

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

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

Question
Answer

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

Read More

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

L-lactate assay kit

Good Good 4/5 (Ease of Use)
Abreviews
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.

Mr. Christian Marx

Verified customer

投稿 Jul 23 2013

Answer



Yes, you can use the syringe. Also use vortexing and sonication alongside it for efficient homogenization. Check the homogenization efficiency under the microscope.

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.

Read More

1-10 of 62 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

登録