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.
Sample Typle |
Plasma,Serum,Cell culture supernatant, other body fluid |
Components |
|
Method |
Colorimetric method at 570nm |
Standard Curve |
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Sensitivity |
2µM-100µM |
Reaction Time |
30 minutes |
Applications |
For biological research: |
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. 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. |
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. |
3.Reaction Solution Preparation |
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. |
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. 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.
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.