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Lactate Colorimetric/Fluorometric Assay Kit

Details for Product No. ABIN411682, Supplier: Log in to see
Antigen
Reactivity
Chemical
4
1
1
1
Detection Range
0.001-10 mM
Minimum Detection Limit
0.001 mM
Application
Biochemical Assay (BCA)
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Purpose In the Lactate Assay Kit, lactate specifically reacts with a enzyme mix to generate a product, which interacts with lactate probe to produce color (570 nm) and fluorescence (at Ex/Em = 535/587 nm). The kit provides a convenient means for detecting L(+)-Lactate in biological samples such as in blood circulation, in cells, in culture mediums, in fermentation mediums, etc. There is no need of pretreatment or purification of samples. The kit can detect 0.001-10 mM of various Lactate samples.
Sample Type Cell and Tissue Culture Supernatant, Plasma, Serum, Tissue Samples, Biological Fluids
Detection Method Fluorometric, Colorimetric
Specificity In the Lactate Assay Kit, lactate specifically reacts with a enzyme mix to generate a product, which interacts with lactate probe to produce color (at lambda = 570 nm) and fluorescence (at Ex/Em = 535/587 nm ). The kit provides a convenient means for detecting L(+)-Lactate in biological samples such as in blood circulation, in cells, in culture mediums, in fermentation mediums, etc. There is no need of pretreatment or purification of samples.
Characteristics Lactate Assay Kit: Colorimetric & Fluorometric Assay to Measure L(+)-Lactate in a variety of Biological Samples such as Serum, Blood, Cells, Culture mediums, Fermentation mediums, etc. within 40 min. Rapid, Simple & Sensitive.
Components Lactate Assay Buffer
Lactate Probe (in DMSO)
Lactate Enzyme Mix
L(+)-Lactate Standard (100 nmol/μl)
Target Type Chemical
Background Abnormal high concentration of lactate has been related to disease states such as diabetes and lactate acidosis, etc. L(+)-Lactate is the major stereo-isomer of lactate 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 specifically reacts with a enzyme mix to generate a product, which interacts with lactate probe to produce color (570 nm) and fluorescence (at Ex/Em = 535/587 nm). The kit provides a convenient means for detecting L(+)-Lactate in biological samples such as in blood circulation, in cells, in culture mediums, in fermentation mediums, etc. There is no need of pretreatment or purification of samples. The kit can detect 0.001-10 mM of various Lactate samples.
Application Notes The kit can detect 0.001-10 mM of various Lactate samples.
Comment

Further details regarding sample type: Cell and tissue culture supernatants, urine, plasma, serum, as well as many other biological fluids

Assay Time < 1 h
Protocol 1. Standard Curve Preparations: For the colorimetric assay, d ilute the Lactate Standard (MW 90.08) to 1 nM/µL 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. For fluorometric assay, dilute the Lactate Standard to 0.1 nM/µL by adding 10 µL of the Lactate Acid to 990 µL of Lactate Assay Buffer, mix well. Then take 20 µL into 180 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, 0.2, 0.4, 0.6, 0.8, 1.0 nM/well of the Lactate Standard.
2. Sample Preparations: Prepare test samples in 50 µL/well with Lactate Assay Buffer in a 96-well plate. If using serum sample, serum (0.5-10 µL/assay, serum contains approx. 0.6 nM/µL lactate) can be directly diluted in the Lactate Assay Buffer. We suggest using several doses of your sample to ensure the readings are within standard curve range. Note: Lactate Dehydrogenase (LDH) may degrade lactate. Therefore, the samples contain LDH (such as culture medium or tissue lysate) should be kept -80C for storage, or filter samples through 10 Kd molecular weight spin filter.
3. Reaction Mix Preparation: Mix enough 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 Probe 2 µL 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, protect from light.
6. Measure O.D. 570 nm for colorimetric assay or fluorescence at Ex/Em = 535/590 nm in a microplate reader. If the background is too high in the fluorometric assay, 1/10 volume of probe may be used, which will decrease the background significantly.
7. Correct background by subtracting the value derived from the 0 lactate control from all sample readings
Restrictions For Research Use only
Storage -20 °C
Expiry Date 12 months
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