EnzyFluo™ L-Lactate Assay Kit
Datasheet PDF  MSDS/Bulk Order


Application
For quantitative determination of L-lactate (L-lactic acid) and evaluation of drug effects on its metabolism.

Key Features
Sensitive and accurate. Detection limit of 1 μM and linearity up to 50 μM L-lactate in 96-well plate assay.

Convenient. The procedure involves adding a single working reagent, and reading the fluorescence after 60 min. Room temperature assay.

High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.

Method
FL530/585 nm

Samples
Serum, plasma, cell culture media etc

Species
All

Procedure
60 min

Size
100 tests

Detection Limit
1 μM

Shelf Life
6 months

More Details
LACTATE is generated by lactate dehydrogenase (LDH) under hypoxic or anaerobic conditions. Monitoring lactate levels is, therefore, a good indicator of the balance between tissue oxygen demand and utilization and is useful when studying cellular and animal physiology. Simple, direct and automation-ready procedures for measuring lactate concentration are very desirable. BioAssay Systems’ EnzyFluo™ lactate assay kit is based on lactate dehydrogenase catalyzed oxidation of lactate, in which the formed NADH reduces a probe into a highly fluorescent product. The fluorescence intensity of this product, measured at λex/em = 530/585 nm, is proportional to the lactate concentration in the sample.

1. Does heparin, EDTA or citric acid interfere with assay?



EDTA and Heparin do not interfere. 10 mM Citric acid lowers signal of 2 mM lactate by 15%.

2. Can this assay be used to determine lactate in cell culture supernatants that contain phenol red?



Yes, lactate assays can be performed in culture media that contains phenol red. We recommend diluting the standard in the control medium (i.e. that does not contain lactate). The detection range is 74 µM to 1 mM. If the lactate concentration in a sample is higher than 1 mM, dilute sample in the medium and repeat assays. Multiply the results by the dilution factor. The detection limit is around 74 µM. Follow the following table for standard dilutions.

3. Can this kit be used to measure lactic acid concentrations in cells?



Yes. To determine intracellular lactic acid concentrations, harvest cells (0.1-2 million cells per assay) in a 1.5mL tube, centrifuge 1-2 min at 3,000 rpm on a table centrifuge. Remove culture medium, wash cells quickly with cold PBS. Immediately remove any PBS. Prepare Working Reagent. Add 120 µL WR directly to the cell pellet and 80uL WR to standards in 96-wells. Vortex cell sample tube 1 min. Transfer 100 µL supernatant to 96-well. Read OD565nm kinetics on a plate reader.

Notes: 1. cell number. It is prudent to run several doses of cells, e.g. 0.1, 1, 2x106 cells to determine optimal cell number to be used in subsequent assays.
2. Working Reagent should contain a lysis reagent. BioAssay Systems will supply this reagent upon request.

4. How to prepare tissue homogenates for use in the assay?



For tissue sample preparation, we recommend the following:

1. Homogenize tissue in phosphate buffered saline. This can be done using a homogenizer (e.g. the Dounce type) or a grinder. If you use a lysis buffer, the buffer should not contain ascorbic acid, SDS, sodium azide, NP-40 and Tween 20.

2. Centrifuge to pellet any debris. This can be done at room temperature in a table centrifuge (e.g. 5 min at 14,000 rpm). The supernatant should be clear.

3. Use the supernatant for lactate assays.

For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions
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EnzyFluo™ L-Lactate Assay Kit
Catalog No: EFLLC-100
Price: $329    Qty:
For orders of 10 or more kits, please contact us at 1-510-782-9988 x 1 for best pricing.

Shipping: On Ice
Shipment: Same Day for order by 2pm PST
Delivery: 1-2 days (US), 3-6 days (Intl);
Storage: -20°C



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