L-Lactate Assay Kit I

Lactate dehydrogenase converts lactate and NAD+ into pyruvate and NADH. The Lactate Assay Kit is based on the reduction of the terazolium salt INT in a NADH-coupled enzyme reaction to formazan, which exhibits an absorbance maximum at 490 nm. The intensity of the absorbance is proportional to the lactate concentration. Using a set of lactate standards, the assay can measure the concentration of lactate in samples (plasma, medium, or others) in a quantitative manner. The kit is stable until the expiration date under proper storage and handling conditions. The product is for research only.

Sku #Product NameProduct SizePriceQTY
120001100A L-Lactate Assay Kit I(5ml) 100 Assays- Assay Solution ONLY (5ml)100 assays- ASSAY SOLUTION ONLY $98.00 USD
1200011002 L-Lactate Assay Kit I (5ml)-100 assays (5ml)-100 assays $108.00 USD
120001100P L-Lactate Assay Kit I(5ml)- 100 Assays w/Reader Plate (1x) (5ml)100 assays w/Reader Plate (1x) $120.00 USD
120001200A L-Lactate Assay Kit I(10ml) 200 Assays- Assay Solution ONLY (10ml)200 assays- ASSAY SOLUTION ONLY $146.00 USD
1200012002 L-Lactate Assay Kit I (10ml)-200 assays (10ml)-200 assays $156.00 USD
120001200P L-Lactate Assay Kit I (10ml)-200 Assays w/ Reader Plate(2x) (10lm)200 assays w/Reader Plate (2x) $180.00 USD
120001400A L-Lactate Assay Kit I (20ml)400 assays- ASSAY SOLUTION ONLY (20ml)400 assays- ASSAY SOLUTION ONLY $267.00 USD
1200014002 L-Lactate Assay Kit I (20ml)-400 assays (20ml)-400 assays $277.00 USD
120001400P L-Lactate Assay Kit I (20ml)-400 Assays w/ Reader Plate(4x) (20ml)400 assays w/Reader Plate (4x) $326.00 USD

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Sample Typle

Plasma,Serum,Cell culture supernatant, other body fluid



Colorimetric method at 490nm

Standard Curve



Reaction Time

30 minutes


For biological research:
L-Lactate measurement in biological samples
For drug/pharm research:
Drug influence on L-Lactate metabolism




Materials needed but not supplied

A plate reader capable of measuring absorbance between 470-490nm

Adjustable pipettes and a repeat pipettor

A source of pure water; glass distilled water of HPLC-grade water is sufficient

0.5M Acetic Acid

Clear flat bottom 96-well plates if not included in the kit purchased

Shipping Icepacks

Reagent Preparation

Note: All reagents are frozen. SWe recommend you spin small vials before opening.

1. L-Lactate Standards
The vial contains 1000µl of 3mM L-Lactate Standard. The standard must be equilibrated to room temperature before use. 1ml of the standard is enough for making 3 standard curves if assayed in duplicate. Store at -80?C.

2. L-Lactate Assay Solution
The solution contains enzymes that are light sensitive. The solution must be thawed on ice before use.Best to aliquot the amount needed and use it all to prevent thawing/freezing cycles.Freeze and store aliquots at -80 oC


1.Sample Preparation

Serum/Plasma/other body fluid/cell culture supernatant
Serum, Plasma, other body fluid, or cell culture supernatant can be measured directly by a series of dilutions of the sample (1/2; 1/4; 1/8; .…..) to ensure the readings are within the standard curve.
Your samples can be diluted with dH2O.
Note: if samples (like hemolyzed serum/plasma or cell culture medium which contain FBS) contain high level of lactate dehydrogenase capable of converting lactate to pyruvate, it is important for the samples to be deproteinated.

Solid Samples
Solid samples, such as tissues, can be first homogenized and extracted with ethanol (80%) with a tissues/Ethanol ratio of 1:8 (1 hr at 4°C) followed by centrifugation at 10,000xg. The clear supernatants then can be measured directly by a series of dilutions of the sample (1/2; 1/4; 1/8; .…..) to ensure the readings are within the standard curve. Your samples can be diluted with dH2O.

Adding Samples
Add 50μl of samples to each well.
We recommend that samples be assayed in duplicate.

If prepared samples are not assayed the same day, store the samples at -80?C. If samples need to be deproteinated, make sure to deproteinize the samples prior to storing in the freezer. The deproteinated samples will be stable for one month while stored at -80?C. For frozen samples, dilutions of samples must be done right before assaying.

2.Standard Curve Preparation
We recommend that L-Lactate Standards be assayed in duplicate. A standard curve has to be run in each assay.
Add 50µl, 40µl, 30µl, 20µl, 10µl, 5µl, 1 µl, and 0µl of L-lactate Standard to each well. Then adjust volume to 50μl/well with dH2O.

3.Perform the assay

a) Add 50ul of L-Lactate assay solution to each well containing L-Lactate standards and test samples.
b) Incubate for 30mins at 37°C incubator. Note: Please do not use CO2 incubator.
c) Stop the reaction by adding 50 μl of 0.5 M acetic acid per well followed by brief gentle agitation. Note: Eliminate any air bubbles present in the wells using a needle prior to measurement.
d) Measure the absorbance at 490nM using a microplate reader.


a) Average the OD490 nm values of replicate wells of each L-Lactate standard, test samples, and blank. In order to get the corrected absorbance, subtract the average OD490 nm value of the blank (L-Lactate Standard #8) from the average OD490 nm values from all standards and samples.

b) Make a standard curve by plotting OD490 nm values from each L-Lactate standards as a function of L-Lactate concentration. This can be done with excel spreadsheet. Calculate the value of L-Lactate in samples using the equation obtained from the linear regression of the standard curve.

L-Lactate(µM)= [ (Corrected absorbance)-(y-intercept)]/slope

Material Safety Data Sheet

Date:Oct 26,2015


1.1 Product identifier

Product name: L-Lactate Assay Kit I

Catalog No: 1200011002,120001100A,120001100P,1200012002,120001200A,12001200P,1200014002,120001400A,120001400P

1.2 Relevant identified uses of the substance or mixture and uses advised against

For research use only.

1.3 Details of the supplier of the safety data sheet

Company:Eton Bioscience Inc




1.4 Emergency telephone number



2.1 Classification of the substance or mixture

Not a hazardous substance or mixture

2.2 GHS Label elements, including precautionary statements

Not a hazardous substance or mixture

2.3 Other hazards



3.1 Substances

L-lactate Assay Solution











L-Lactate Assay Standard



Sodium L-Lactate







4.1 Description of first aid measures

Eye contact

Check for and remove contact lenses and flush with copious amounts of water; assure adequate flushing by separating the eyelids with fingers; call a physician

Skin Contact

Flush with copious amounts of water; remove contaminated clothing and shoes; call a physician


Remove to fresh air. If not breathing give artificial respiration ;call a physician


If swallowed, never give anything by mouth to an unconscious person. Rinse mouth with water ;call a physician

4.2 Most important symptoms and effects, both acute and delayed

No information available

4.3 Indication of any immediate medical attention and special treatment needed.

No information available


5.1 Extinguishing media

Suitable extinguishing media

Water spray, alcohol-resistant foam, dry chemical or carbon dioxide

5.2 Special hazards arising from the substance or mixture

No information available

5.3 Advice for firefighters

Wear self-contained breathing apparatus and protective clothing to prevent contact with skin and eyes.


6.1 Personal precautions,protective equipment and emergency procedures

Use full personal protective equipment. Evacuate personnel to safe areas. Avoid breathing vapors,mist or gas.

6.2 Environmental precautions

Prevent further leakage or spillage. Prevent product from entering drain.

6.3 Methods and material for containment and cleaning up

Keep in suitable, closed containers for disposal


7.1 Precautions for safe handling

Avoid skin/eye contact. Use protective equipment as needed. Wash contaminated clothing before reuse.

7.2 Conditions for safe storage,including any incompatibilities

Keep container tightly closed. Store at -80?C.

7.3 Specific end use(s)

No data available.


8.1 Control Parameters

This product contains no hazardous materials with occupational exposure limits.

8.2 Exposure controls

Engineering controls

Provide shower and eye wash station

Personal protective equipment

Eye Protection

Wear safety goggles

Skin Protection

Wear protective clothing

Hand Protection

Wear protective gloves

Respiratory protection

Wear NIOSH/MSHA approved respirators


9.1 Information on basic physical and chemical properties

L-Lactate Assay Solution


Liquid, yellowish



Melting point

No data available

Freezing point

No data available

Boiling point

No data available


No data available

Water solubility


L-Lactate Standard


Liquid, colorless



Melting point

No data available

Freezing point

No data available

Boiling point

No data available


No data available

Water solubility


9.2 Other safety information

No data available.



No data available

10.2Chemical stability

Stable under recommended storage conditions.

10.3Possibility of hazardous reactions

No data available.

10.4Conditions to avoid

No data available

10.5Incompatible materials

Strong oxidizing agents

10.6Hazardous decomposition products

Hazardous decomposition products formed under fire conditions: carbon oxides


11.1Information on toxicological effects

Toxicological effects on this product have not been thoroughly studied.



Avoid release into environment

12.2Persistence and degradability

No data available

12.3.Bioaccumulative potential

No data available

12.4Mobility in soil

No data available

12.5Results of PBT and vPvB assessment

No data available

12.6Other adverse effects

No data available


13.1Waste treatment methods

Dispose in accordance with local, state, and federal regulations.



Proper shipping name: none
Non-Hazardous for transport: this substance is considered to be non-hazardous for transport

Proper shipping name: none
Non-Hazardous for transport: this substance is considered to be non-hazardous for transport


Proper shipping name: none
Non-Hazardous for transport: this substance is considered to be non-hazardous for transport


EU Risk and Safety phrases:

R36/37/38: irritating to eyes/respiratory system/skin


The information above is believed to be accurate and represents the best information currently available to us. However, we make no warranty of merchantability or any other warranty, express or implied, with respect to such information. Eton Bioscience Inc. shall not be held liable for any damages or other consequences resulting from handling or from contact with the above product.

Q. What type of medium should I use for making cultured cells for Lactate assay?
A. Please use phenol red free medium. Please do not use phenol red medium since phenol red would affectabsorbance readings.

Q. What enzymes is used in Lactate Assay?
A. It is LDH.

Q. Can you tell me the compositions of your Lactate Assay?
A. Unfortunately it is proprietary.

Q. How do I know whether I need to use L-Lactate or D-Lactate assay kit?
A. L-Lactate assay kits are commonly used for measuring lactate level in animal cells.
D-lactate assay kits are commonly used for measuring lactate level in bacteria cells.

Q. Do I need to make a standard curve everytime?
A. Yes, it is necessary since you calculate your sample concentration based on the standard curve. Please do not usethe representative Lactate standard curve in the protocol to calculate concentrations of your samples.

Q.Can I use fluorescence spectroscopy to measure readings for Lactate assay kits?
A. No, the kit does not work with a fluorescence reader since the kit employs colorimetric assays.

Q. How do I measure Lactate from cultured cells?
A. You can grow cells in phenol red-free medium, then you can extract cells with 80% ethanol.

Q. Can I use frozen samples?
A. Although it is better to use fresh samples for assays, you canuse frozen samples. If you are not using frozen samples in once, please make aliquots of your samples before you put them in a freezer to prevent from degrading samples from repeated freeze-thaw cycles.
Q. My sample formed preciptation after adding acetic acid. How can I prevent this?
A. You can still measure readings without adding acetic acid.
Q. The protocol recommends measuring the absorbance at 490nm. Is 485nm close enough?
A. It is ok as long as you set up the absorbance between 470 and 490nm.
Q. Will L-Lactate assay measure D-Lactate?
A. No, it wont.
Q. Can I use a transparent wall 96-well plate with Lactate Kit?
A. Yes, you can.

Q. Can Ethanol interfere with the enzyme?

A. The enzyme can tolerate up to 20% of ethanol without affecting it’s enzymatic activity with L-Lactate.

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