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Lactate Assay Kit
Biochemical Assay (BCA)
|9 references available|
|Price||423.50 $ Plus shipping costs $45.00|
|Availability||Will be delivered in 2 to 3 Business Days|
|Sample Type||Cell and Tissue Culture Supernatant, Urine, Plasma, Serum, Biological Fluids|
|Components||Lactate Assay Buffer, Lactate Probe (in DMSO), Lactate Enzyme Mix, L(+)-Lactate Standard (100 nmol/µl)|
|Description||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 (at λ = 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.|
|Protocol||1. Standard Curve Preparations: For the colorimetric assay, d ilute the Lactate Standard (MW 90.08) to 1 nmol/ μ l by adding 10 μ l of the Lactate Standard to 990 μ l of Lactate Assay Buffer, mix well. Add 0, 2, 4, 6, 8, 10 μ l into each well individually. Adjust volume to 50 μ l/well with Lactate Assay Buffer to generate 0, 2, 4, 6, 8, 10 nmol/well of the L(+)- Lactate Standard. For fluorometric assay, dilute the Lactate Standard to 0.1 nmol/ μ l by adding 10 μ l of the Lactate Acid to 990 μ l of Lactate Assay Buffer, mix well. Then take 20 μ l into 180 μ l of Lactate Assay Buffer. Mix well. Add 0, 2, 4, 6, 8, 10 μ l into each well individually. Adjust volume to 50 μ l/well with Lactate Assay Buffer to generate 0, 0.2, 0.4, 0.6, 0.8, 1.0 nmol/well of the Lactate Standard. 2. Sample Preparations: Prepare test samples in 50 μ l/well with Lactate Assay Buffer in a 96-well plate. If using serum sample, serum (0.5-10 μ l/assay, serum contains ~0.6 nmol/ μ l 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 (BioVision, Cat 1997). 3. Reaction Mix Preparation: Mix enough reagent for the number of assays performed: For each well, prepare a total 50 μ l Reaction Mix containing the following components. Mix well before use. 46 μ l Lactate Assay Buffer 2 μ l Probe 2 μ l Enzyme Mix 4. Add 50 μ l 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 (|
|Expiry Date||12 months|
|Restrictions||For Research Use only|
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Further details: Lactate Assay Extracellular lactate measurement was performed using the Lactate Assay Kit
Rey, Luo, Shimoda et al.: "Metabolic reprogramming by HIF-1 promotes the survival of bone marrow-derived angiogenic cells in ischemic tissue." in: Blood, Vol. 117, Issue 18, pp. 4988-98, 2011 (PubMed).
Zhao, Liu, Liu et al.: "Overcoming Trastuzumab Resistance in Breast Cancer by Targeting Dysregulated Glucose Metabolism." in: Cancer research, Vol. 71, Issue 13, pp. 4585-4597, 2011 (PubMed).
Varum, Rodrigues, Moura et al.: "Energy metabolism in human pluripotent stem cells and their differentiated counterparts." in: PLoS ONE, Vol. 6, Issue 6, pp. e20914, 2011 (PubMed).
Nakano, Tsuji, Miki et al.: "Glycolysis Inhibition Inactivates ABC Transporters to Restore Drug Sensitivity in Malignant Cells." in: PLoS ONE, Vol. 6, Issue 11, pp. e27222, 2011 (PubMed).
Lee, Yi, Ku et al.: "A pumpless perfusion cell culture cap with two parallel channel layers keeping the flow rate constant." in: Biotechnology progress, 2012 (PubMed). Further details: For concentration measurement of lactate and ammonia, a lactate assay kit and an ammonia assay kit are used, respectively.