Powdered GC muscles (10–20 mg per assay) were sonicated in the appropriate assay buffer (Q700 ice-water bath sonicator, QSONICA) for six cycles of amplitude 35 for 30 sec followed by 1 min on ice, centrifuged at 10,000 × g for 10 min at 4 °C and the lysates were subjected to the following assays: measurement of the metabolites and citrate synthase (CS) activity. Lactate Colorimetric Assay (Biovision, #K607-100), Glycogen Colorimetric Assay (Biovision, #K648-100), and Citrate Synthase Activity Colorimetric Assay (Biovision, #K318-100) were performed according to the manufacturers’ instructions.
Lactate colorimetric assay
Manufactured by Abcam
The Lactate Colorimetric Assay is a biochemical test used to quantify the concentration of lactate in a sample. The assay utilizes an enzymatic reaction that produces a colored product, which can be measured using a spectrophotometer. The intensity of the color is proportional to the amount of lactate present in the sample.
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Lab products found in correlation
Glucose uptake colorimetric assay, by Abcam (2 mentions)
Glycogen colorimetric assay, by Abcam (1 mentions)
Q700 ice water bath sonicator, by Qsonica (1 mentions)
Pierce ldh cytotoxicity assay kit, by Thermo Fisher Scientific (1 mentions)
Gem premier 4000, by Werfen (1 mentions)
White 96 well plate, by Corning (1 mentions)
Celltiter glo luminescent cell viability assay, by Promega (1 mentions)
4 protocols using lactate colorimetric assay
1
Muscle Metabolite and Enzyme Assays
2
Metabolic Profiling of Cancer Cells
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The glucose uptake colorimetric assay, lactate colorimetric assay and ATP colorimetric assay kits (Biovision, Inc.) were used to examine glucose consumption and lactate and ATP production in A549 and H1299 cells, according to the manufacturer's protocols.
3
Glucose and Lactate Assay for Cancer Cells
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The glucose uptake colorimetric assay (Biovision) and the lactate colorimetric assay (Biovision) kits were used to detect glucose consumption and lactate production in GC cells, according to the manufacturer’s instructions.
4
Metabolic Analysis of Cell Cultures
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The level of lactate and glucose in culture medium was measured by the nuclear magnetic resonance (NMR) analysis as described below, or by blood gas analyzer GEM Premier 4000 (Instrumentation Laboratory, Bedford, MA). Additionally, the lactate concentration was measured by Lactate Colorimetric Assay (Biovision, Milpitas, CA) following the manufacturer instructions. The enzymatic activity of intracellular lactate dehydrogenase (LDH) was measured in 96-well plates by using the Pierce LDH Cytotoxicity Assay Kit (Thermoscientific, Rockford, IL) following the manufacturer instructions. For ATP, the cells were cultured in 96-well white plates (Corning, NY), and the ATP level was detected by CellTiter-Glo ® Luminescent Cell Viability-assay (Promega, Madison, WI) measuring ATP-dependent bioluminescence on a Victor plate reader.
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