To dissect mechanism of anti-tumor response with indicated drug combinations, quantification of lactate was performed using colorimetry kit (Biovision; K627–100) on both ex vivo and in vitro CM, as described above.
K627 100
Manufactured by Abcam
Sourced in United States
K627-100 is a lab equipment product. It is a 100-gram pack of a chemical reagent.
Automatically generated - may contain errors
Lab products found in correlation
L lactate assay kit, by Abcam (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
D glucose, by Merck Group (2 mentions)
Bca protein assay, by Thermo Fisher Scientific (2 mentions)
Colorimetry kit, by Abcam (1 mentions)
K666 100, by Abcam (1 mentions)
Glucose colorimetric assay kit 2, by Abcam (1 mentions)
K676 100, by Abcam (1 mentions)
K606 100, by Abcam (1 mentions)
Enliten atp assay system, by Promega (1 mentions)
Lactate colorimetric assay kit, by Abcam (1 mentions)
K686 100, by Abcam (1 mentions)
Seahorse xf96 extracellular flux analyzer, by Agilent Technologies (1 mentions)
10 protocols using k627 100
1
Quantifying Tumor Microenvironment Metabolomics
2
Quantifying Cellular Metabolism Rates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
3
Glucose and Lactate Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Glucose levels were determined using a commercial glucose assay kit, Glucose Colorimetric assay kit II (K686-100, Bio Vision, Milpitas, CA, USA). Glucose uptake was calculated by deducting the detected glucose concentration in the medium from the original glucose concentration. Lactate levels were determined using a lactate colorimetric assay kit (K627-100, Biovision, Milpitas, CA, USA) in accordance with the manufacturer’s instructions. Considering that the cell number in every sample may be different, all the concentrations of glucose or lactate production were finally normalized to the total cell protein concentration. The protein concentration was measured by the BCA protein assay (Cat. 23225, Thermo Fisher Scientific, Waltham, MA, USA).
4
Colorimetric Quantification of Glucose Uptake and Lactate Production
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
5
Lactate Release in Oligodendrocyte Progenitor Cells
6
Lactate Release in Oligodendrocyte Progenitor Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine the lactate release, OPCs were seeded into 24-well plates at a density of 5 × 104 cells/well and cultured until the desired stage in appropriate medium. The culture medium was first changed to 250 μL of glucose-free DMEM medium (GIBCO Cat# 11966025) to starve the cells for 1 h, then 25 mM D-glucose (Sigma Cat# G8270) was added into the medium to initiate glycolysis. For the hM4Di OPCs, 5 μM CNO was added at the beginning of glucose starvation. The medium was collected 1 h after glucose treatment and treated with two volumes of acetone immediately to precipitate protein. Samples were then lyophilized to remove acetone and re-dissolved in sterilized deionized water. For tissue lactate content detection, mouse brains were dissected, and the hypothalamus was snapfrozen in liquid nitrogen within 20 s. The isolated hypothalamus samples were weighed and homogenized in 50% methanol to precipitate protein. After centrifuging at 4°C, the homogenate was lyophilized and re-dissolved by sterilized deionized water. Lactate levels were measured using the L-lactate assay kit according to the manufacturer’s protocol (Biovision Cat# K627–100).
7
Metabolic Profiling of Hepatocellular Carcinoma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hepatocellular carcinoma cells are handled in accordance with the needs of the experiment. Using a Lactate Colorimetric Assay kit II, the buildup of lactate in the medium was found (K627-100, BioVision, USA). Utilizing a glucose colorimetric assay kit, the quantity of glucose was determined (K606-100, Bio Vision, USA). The ENLITEN® ATP Assay System (Promega) was used to quantify ATP release in order to examine the ATP in supernatants.
8
Measuring Lactate Production in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded in a 24-well plate in the growth medium containing 10% FBS under standard culture condition. Once cells reached 90% confluence, the growth media were removed, and cells were washed twice in PBS and incubated in 2% serum and phenol-red free medium for 24 h. Then, the media were collected for lactate production assay. L-(+)-Lactate was measured with a colorimetric kit (BioVision, K627-100) according to the manufacturer’s instructions. Absorbance (optical density 450 nm) for each sample was background corrected with the culture medium (2% FBS) collected from the well without growing cells. Final data of the lactate production rates were normalized to the protein amount per well.
Lactate and glucose concentration measurement was also done by a YSI machine and followed their protocol (YSI 2900 multianalyte system [YSI, Yellow Springs, OH]).
Lactate and glucose concentration measurement was also done by a YSI machine and followed their protocol (YSI 2900 multianalyte system [YSI, Yellow Springs, OH]).
9
Glycolysis Efficiency Measurement in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Glucose uptake and lactate production were calculated by detecting the glucose content of the medium to evaluate the glycolysis efficiency of the cells. The glucose level was measured using a glucose colorimetric assay kit II (K686-100, BioVision, Milpitas, CA, USA). A lactic acid colorimetric assay kit (K627-100, BioVision) was used to determine the lactic acid level. All testing procedures were carried out in accordance with the manufacturer’s instructions. The Seahorse Extracellular Flux Analyzer XF96 (Seahorse Bioscience, www.seahorsebio.com ) was used to monitor in vitro cells metabolic alternations, according to the manufacturer’s instructions. For detection of the real-time glycolytic rate, cells were incubated with unbuffered medium followed by a sequential injection of 10 mM glucose, 1 µM oligomycin (OM), and 80 mM 2-deoxyglucose (2-DG). Oligomycin and 2-deoxyglucose were purchased from Topscience (shanghai, China). Considering that the number of cells in each sample may be different, the concentrations of glucose or lactic acid and ECAR measurements were normalized to the cell number.
10
Lactate Production Assay in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded in a 24-well plate in the growth medium containing 10% FBS under standard culture condition. Once cells reach 90% confluence, the growth media were removed and cells were washed twice in PBS and incubated in 2% serum and phenol-red free medium for 24 h. Then, the media were collected for lactate production assay. L (+)-Lactate was measured with a colorimetric kit (BioVision, K627-100) according to the manufacturer's instructions. Absorbance (OD 450 nm) for each sample was background corrected with the culture medium (2% FBS) collected from the well without growing cells. Final data of the lactate production rates were normalized to the protein amount per well. Lactate and Glucose concentration measurement was also done by YSI machine followed their protocol (YSI 2900 multi-analyte system (YSI, Yellow springs OH)).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!