L-Lactate Assay Kit II

Lactic acid, a major stereoisomer of lactate formed in human intermediary metabolism, has often been used as a physiological indicator for stress. Its determination in serum (normal range in blood 0.5-2.0 mM) is very important in the diagnosis and medical management of various diseases, such as tissue hypoxia, diabetes, circulatory failure and hematological disorders. Unlike a typical lactic acid-LDH assay, this new lactic acid assay kit provides a simple, reliable method for quantifying lactic acid in biological samples such as blood (serum and plasma), culture and fermentation media, etc. In the assay, lactic acid is oxidized by enzyme reactions to yield color product, which can be measured at 570 nm for colorimetric assay, And the color intensity is proportional to lactic acid concentrations, therefore the sample lactic acid concentration can be accurately calculated based on the lactic acid standards.

Sku# Product Name Product Size Price QTY

Sku#

120005200P

Product Name

L-Lactate Assay Kit II-200 Assays w/Reader Plate(2x)

Product Size

200 Assays w/Reader Plate (2x)

Price

$501.60 USD

QTY

Sku#

1200052002

Product Name

L-Lactate Assay Kit II 200 Assays

Product Size

200 Assays

Price

$475.20 USD

QTY

Sku#

120005100P

Product Name

L-Lactate Assay Kit II-100 Assays w/Reader Plate(1x)

Product Size

100 Assays w/Reader Plate (1x)

Price

$286.00 USD

QTY

Sku#

1200051002

Product Name

L-Lactate Assay Kit II 100 Assays

Product Size

100 Assays

Price

$264.00 USD

QTY

Add to Cart

Sample Typle

Plasma,Serum,Cell culture supernatant, other body fluid

Components

Method

Colorimetric method at 570nm

Standard Curve

Sensitivity

2µM-100µM

Reaction Time

30 minutes

Applications

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

Storage

-80°C

Notes

Material needed but not supplied

A plate reader capable of measuring absorbance at 570nm

Adjustable pipettes and a repeat pipettor

Distilled water(milliQ or HPLC-grade)

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

Shipping Icepacks

Reagent Preparation

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

1. L-Lactate Assay Buffer(10x)
Note: Please don’t do dilution in the providedL-Lactate Assay Buffer bottle itself.

The bottle contains 10ml of 10x L-Lactate Assay Buffer. The assay buffer must be equilibrated to room temperature before use. Please make appropriate amount of 1xL-Lactate Assay Buffer for the assay, for example, by diluting 1ml of 10x L-Lactate Assay Buffer with 9 ml of distilled water. Prepare additional diluted assay buffer if needed. Store at -80?C.
2. L-Lactate Standard (10x)
Note: Please don’t do dilution in the provided L-Lactate standard vial itself.

The vial contains 100µl of 1mM L-Lactate Standard. The standard must be equilibrated to room temperature before use. Dilute 100µl of 1mM L-Lactate Standard with 900µl of diluted L-Lactate Assay Buffer to prepare a 100μM L-Lactate Standard. 1ml of diluted standard is enough for making 3 standard curves if assayed in duplicate. Store at -80?C.

3. Assay Probe
The vial contains 45μl of Assay Probe. It is sufficient for 100 wells. The assay probe must be equilibrated to room temperature before use.Protect from light. Store at -80?C.

4. L-Lactate Enzyme Mix
The vial contains 55μl of L-Lactate Enzyme Mix. The enzyme mix must be thawed on ice before use. It is sufficient for 100 wells. Store at -80?C.

Protocol

1.Sample Preparation

Blood, 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 range. Your samples can be diluted with dH2O or diluted L-Lactate Assay Buffer.However, keep in mind if samples (such as hemolyzed serum/plasma) contain high level of lactate dehydrogenase capable of converting lactate to pyruvate, it is important for the samples to be deproteinated.

Add 50μl of samples to each well. We suggest that samples be assayed in duplicate.

Note:
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 Prepation

Add 50µl, 40µl, 30µl, 20µl, 10µl, 5µl, 1 µl, and 0µl of diluted L-Lactate Standard to each well, then adjust volume to 50μl/well with diluted L-Lactate Assay Buffer.
We suggest that L-Lactate Standards be assayed in duplicate. A standard curve has to be included with samples in each assay.

3.Reaction Solution Preparation
50μl Reaction Solution is required for each well, and 1ml of Reaction Solution is prepared with the followings:

982µlL-Lactate Assay Buffer(1x)
10µlL-Lactate Assay Enzyme Mix
8 µlAssay Probe

1ml of Reaction Solution is enough for ~20 wells.

Prepare appropriate amount of Reaction Solution for the number of assays to be performed.

Note:Please do not keep any leftover Reaction Solution as it has to be made fresh right before you run assays. If old Reaction Solution is used , assays will not work.

4.Perform the assay

a. Mix the reaction solution well and add 50μl of the reaction solution to each well containing the L-Lactate standards and test samples.

b. Incubate the reaction for 30 minutes in a 37?C incubator.

c. Measure absorbance at 570 nm in a microplate reader.

5. Calculation

a. Average the OD570 nm values of replicate wells of each L-Lactate Standard, test samples, and blank. Then, correct background by subtracting the value derived from the blank from all sample readings.

b. Make a standard curve by plotting OD570 nm values from each L-Lactate Standards as a function of L-Lactate concentration. This can be done with excel spreadsheet.

c. 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:Sept 21,2017

1. PRODUCT AND COMPANY IDENTIFICATION

1.1 Product identifier

Product name: L-Lactate Assay Kit II

Catalog No: 1200051002, 120005100P, 1200052002, 12005200P

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

TollFree:1-800-758-1630

Tel:1-800-758-1630

Fax:1-800-507-2912

1.4 Emergency telephone number

1-800-758-1630

2. HAZARDS IDENTIFICATION

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

None

3. COMPOSITION/INFORMATION ON INGREDIENTS

3.1 Substances

L-Lactate Assay Buffer

L-Lactate Assay Enzyme Mix

Components

CAS#

Components

CAS#

Potassium Phosphate Monobasic

7778-77-0

Potassium Phosphate Monobasic

7778-77-0

Potassium Phosphate Dibasic

7758-11-4

Potassium Phosphate Dibasic

7758-11-4

Water

7732-18-5

Lactate Oxidase

9028-72-2

L-Lactate Assay Standard

Assay Probe

Components

CAS#

Components

CAS#

Sodium L-Lactate

867-56-1

DMSO

67-68-5

Potassium Phosphate Monobasic

7778-77-0

Potassium Phosphate Dibasic

7758-11-4

Water

7732-18-5












4. FIRST AID MEASURES

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

Inhalation

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

Ingestion

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. FIRE FIGHTING MEASURES

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. ACCIDENTAL RELEASE MEASURES

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. HANDLING AND STORAGE

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. EXPOSURE CONTROLS/PERSONAL INFORMATION

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. PHYSICAL AND CHEMICAL PROPERTIES

9.1 Information on basic physical and chemical properties

L-Lactate Assay Buffer

Appearance

Liquid, colorless

pH

7.0

Melting point

No data available

Freezing point

No data available

Boiling point

No data available

Density

No data available

Water solubility

Soluble

L-Lactate Standard

Appearance

Liquid, colorless

pH

7.0

Melting point

No data available

Freezing point

No data available

Boiling point

No data available

Density

No data available

Water solubility

Soluble

L-Lactate Assay Enzyme Mix

Appearance

Liquid, yellow

pH

No data available

Melting point

No data available

Freezing point

No data available

Boiling point

No data available

Density

No data available

Water solubility

Soluble

Assay Probe

Appearance

Liquid, colorless

pH

No data available

Melting point

No data available

Freezing point

No data available

Boiling point

No data available

Density

No data available

Water solubility

Soluble

9.2 Other safety information

No data available.

10. STABILITY AND REACTIVITY

10.1Reactivity

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. TOXICOLOGICAL INFORMATION

11.1Information on toxicological effects

Toxicological effects on this product have not been thoroughly studied.

12. ECOLOGICAL INFORMATION

12.1Toxicity

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. DISPOSAL INFORMATION

13.1Waste treatment methods

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

14. TRANSPORT INFORMATION

DOT:

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

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

ADR/RID

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

15. REGULATORY INFORMATION

EU Risk and Safety phrases:

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

16. OTHER INFORMATION

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.

1. Marbach E. P. and M. H. Weil (1967) Rapid enzymatic measurement of blood lactate and pyruvte. Clin. Chem. 13: 314-320.
2. van Ginneken, V. R. Boot, T. Murk, G. van den Thillart, and P. Balm.2004. Blood plasma substrates and muscle lactic-acid response after exhaustive exercise in common carp and trout: indications for a limited lactate-shuttle. Anim. Biol. Leiden Neth. 54:119–130.
3. Noordally O, Vincent J-L (1999) Evaluation of a new, rapid lactate analyzer in critical care. Intens Care Med 25:508-513.

What enzyme is used in this assay?

It is Lactate Oxidase.

What are the differences between L-Lactate Assay Kit I and L-Lactate Assay Kit II?

Here are some differences between L-Lactate Assay Kit I and L-Lactate Assay Kit II.

L-Lactate Assay Kit I

Standard Curve range: 50μM-3200μM

Enzyme used: L-LDH

Colorimetric reader compatible

Components: Assay Solution and Pre-serial diluted Standards

L-Lactate Assay Kit II

Standard Curve range: 2μM-100μM

Enzyme used: Lactate Oxidase

Colorimetric reader compatible

Components: Assay Buffer,Enzyme mix,Assay Probe,and Assay standard

What are the compositions of this assay?

Unfortunately it is proprietary.

Do I need to make a standard curve every time?

Yes it is necessary since you calculate your sample concentration based on the standard curve. Please do not use the representative Lactate standard curve in the protocol to calculate concentrations of your samples.

How much sample do I need for this assay?

End-users would have to perform an experiment to determine optimal volume of samples whose readings fall within the standard curve range.

Can I use frozen samples?

Although it is better to use fresh samples for assays, you can use 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.

The protocol recommends measuring the absorbance at 570nm. Is 565nm close enough?

It is ok as long as you set up the absorbance between 560 and 580nm.

Why are my standard curve values different than the one shown on the datasheet?

Many factors influence the values such as room temperature, incubation time, handling, etc.

Are trial kits available?

Yes they are available for first time customers, however, you will be responsible for shipping and handling fee. Please contact

for more details.


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