Lactate Colorimetric Assay Kit II Kit
- Detection Method
- Detection Range
- 0.02-10 mM
- Minimum Detection Limit
- 0.02 mM
- Biochemical Assay (BCA)
- Sample Type
- Cell Culture Supernatant, Plasma, Serum, Tissue Samples
- In the Lactate Assay Kit, lactate is oxidized by lactate dehydrogenase to generate a product which interacts with a probe to produce a color (lambda max = 450 nm). The kit detect L(+)-Lactate in biological samples such as in serum or plasma, cells, culture and fermentation media. There is no need for pretreatment or purification of samples. It detects 0.02 mM - 10 mM lactate in various samples.
- Lactate Assay Kit II: Colorimetric Assay to Measure L(+)-Lactate in a variety of Biological Samples such as Serum, Plasma, Cells, Culture & Fermentation mediums, etc. within 40 min. Rapid, Simple & Sensitive.
Lactate Assay Buffer
Lactate Enzyme Mix
Lactate Substrate Mix
L(+)-Lactate Standard (100 mM)
- Application Notes
- Kit detects 0.02 mM - 10 mM lactate in various samples.
Further details regarding sample type: Cell and tissue culture supernatants, urine, plasma and serum, as well as many other biological fluids, monitoring level during fermentation and feeding in protein expression processes
- Assay Time
- < 1 h
1. Standard Curve Preparations : Dilute the Lactate Standard (MW 90.08) to 1 mM by adding 10 µL of the Lactate Standard to 990 µL of Lactate Assay Buffer, mix well. Add 0, 2, 4, 6, 8, 10 µL into each well individually. Adjust volume to 50 µL/well with Lactate Assay Buffer to generate 0, 2, 4, 6, 8, 10 nM/well of the L(+)-Lactate Standard.
2. Sample Preparation: Prepare test samples at 50 µL/well with Lactate Assay Buffer in a 96-well plate. For serum samples, 0.5-10 µL serum can be directly tested (regular serum contains approx. 0.6 nM/µL lactate). We suggest using several doses of your sample to ensure the readings are within the standard curve range. Note: (1) Tissue or cells can be homogenized in the assay buffer. Centrifuge to remove the insoluble materials. The soluble fraction may be assayed directly. (2) NADH or NADPH from cell or tissue extracts generates background for the lactate assay. To remove the NADH or NADPH background, same amount of sample can be tested in the absence of Lactate Enzyme Mix. Then the background readings can be subtracted from the lactate reading. (3) Endogenous Lactate Dehydrogenase (LDH) may degrade lactate. Samples containing LDH (such as culture medium or tissue lysate) should be kept at -80 °C for storage, or filtered through a 10kd mw spin filter to remove all proteins.
3. Reaction Mix Preparation: Mix sufficient reagent for the number of assays performed. For each well, prepare a total 50 µL Reaction Mix containing the following components. Mix well before use: 46 µL Lactate Assay Buffer 2 µL Lactate Substrate Mix 2 µL Lactate Enzyme Mix
4. Add 50 µL of the Reaction Mix to each well containing the Lactate Standard or test samples, mix well.
5. Incubate the reaction for 30 minutes at room temperature.
6. Measure O.D. 450nm in a microplate reader. The color is stable for at least 4 hours.
- Calculation of Results
Calculation: Correct background by subtracting the value derived from the 0 lactate control from all standard and sample readings
- For Research Use only
- -20 °C
- Expiry Date
- 12 months
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- Target Type
- Lactate (CH₃CH(OH)COO-) plays important roles in many biological processes. Abnormally high concentrations of lactate have been related to disease states such as diabetes and lactic acidosis, etc. L(+)-Lactate is the major lactate stereoisomer 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. In the Lactate Assay Kit, lactate is oxidized by lactate dehydrogenase to generate a product which interacts with a probe to produce a color (λmax = 450 nm). The kit detect L(+)-Lactate in biological samples such as in serum or plasma, cells, culture and fermentation media. There is no need for pretreatment or purification of samples. It detects 0.02 mM - 10 mM lactate in various samples.