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Lactate Colorimetric/Fluorometric Assay Kit

based on 113 citations in multiple journalsLactate Colorimetric/Fluorometric Assay Kit1135 5
Catalog #: K607

In stock

$475.00

Product Details

Cat # +Size K607-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The kit can detect 0.001-10 mM of various Lactate samples.
Features & Benefits • Simple procedure; takes ~40 minutes
• Fast and convenient
• Kit contains the necessary reagents for accurate measurement of Lactate colorimetrically and fluorometrically
Kit Components • Lactate Assay Buffer
• Lactate Probe (in DMSO)
• Lactate Enzyme Mix
• L(+)-Lactate Standard (100 nmol/µl)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

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.


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Is there difference between incubation of 37 C and Room Temp?
The difference between room temperature and 37 C is not significant enough to affect the results. The reaction is slower at RT.
What is the explanation for seeing lower readings at higher conc. of sample?
Higher concentration of Lactate does have inhibitory effect that leads to lower readings. So, smaller amount of sample is recommended to generate values that fit in the linear part of the standard curve. When there is lactate overdose, the reaction will appear dark pink and then turns brown.
What kind of medium should be used with this assay?
Medium devoid of Lactate and/ or pyruvate should be used. Medium containing FBS should be deproteinized to remove LDH which can degrade Lactate.
Is it essential to deproteinize samples for this assay?
Yes, it is highly recommended to deproteinize samples/medium to remove enzymes such as LDH which can quickly degrade Lactate.
Can frozen samples be used with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples (-80C) can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods. For cell lysates/tissue homogenates or cell culture media, storing after deproteinizing is recommended.
How many cells should be grown per well so that the medium can then be assayed for lactate?
There are several variables : 1) Cell type and the metabolic sate of the cell 2) Medium glucose level 3) lactate/glucose ratio in the media. Since cell types vary, the number of cells used depends on the cell type and experimental data needs to be generated to predict the level of lactate Vs. time. As time passes, cells will be consuming glucose and produce lactate. As the level of glucose drops and the cell starve, they start consuming lactate. So, the level of lactates goes up with time and eventually it starts decreasing.
Can this assay be normalized with respect to protein concentration?
Since deproteinizing is important for preserving the lactate in the sample, it is difficult to normalize lactate concentration with protein in the sample. However, sample can be added to the deproteinizing spin filters based on total protein.
Can this assay be automated and used with 384-well plates?
Although we have optimized the volumes of reagents based on tests with a 96-well plate format, this assay can be automated and used with 384 well plates. It will be essential to proportionately scale down the reagents.
Will the phenol red in the media affect the assay readout?
Very low amounts of media are used for each sample. This will generate a very low background at the best. Please use only media as a background control and subtract this reading from all sample readings to accommodate for the phenol red.
Can the buffer from K627 be used for this assay?
Since the detection principles for the kits K627 and K607 are different, it is essential to use the buffer provided in the kit for best results.
Does this asay detect D-lactate as well?
No, this assay is specific for L-lactate. If there is a substantial amount of D-lactate in the sample, a parallel background control can be run to exclude any possibility of interference.
Can this kit be measured using 530nm filter?
It is always recommended to use the exact recommended wavelength for the most accurate results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from 570nm.
Ashley R. Maiuri, et al., (2018) Inflammation-induced DNA methylation of DNA polymerase gamma alters the metabolic profile of colon tumors, Cancer Metab, Jul. 2018, 30002826
Nai-Yuan Liu, et al., (2018) A Handheld, Colorimetric Optoelectronic Dynamics Analyzer for Measuring Total Ammonia of Biological Samples, Journal of Translational Engineering in Health and Medicine, Jul. 2018, 30112251
Ana I. Duarte, et al., (2018) Dual Therapy with Liraglutide and Ghrelin Promotes Brain and Peripheral Energy Metabolism in the R6/2 Mouse Model of Huntington’s Disease, Scientific Reports, Jun. 2018, 29895889
Paige M. Kulling, et al., (2018) Dysregulation of the IFN-γ-STAT1 signaling pathway in a cell line model of large granular lymphocyte leukemia, Plos one, Feb. 2018, 29474442
Gu Z et al., (2017) NEK2 Promotes Aerobic Glycolysis in Multiple Myeloma Through Regulating Splicing of Pyruvate Kinase, J Hematol Oncol, 2017, 13;10(1):17
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