Oxygen consumption was measured using an extracellular oxygen consumption kit (Abcam) following the manufacturer’s instructions. Briefly, overnight cultures were synchronized and grown for 3 h in M9 media supplemented with 1% casamino acids. These were then used to standardized cultures to OD600 0.1 in the same media with or without 0.2% glucose. Standardized cultures (150 μL) were mixed with 10 μL of oxygen consumption reagent in a black 96-well plate and sealed with 100 μL mineral oil. Extracellular oxygen consumption was monitored using a Cytation 5 plate reader (BioTek) for 2 h in 1 min 40s intervals (excitation = 380 nm; emission = 645 nm) at 37°C. The rate of oxygen consumption was calculated by determining slope values for each sample after 30 min. Results are the average of three biological replicates.
Extracellular oxygen consumption kit
Manufactured by Abcam
Sourced in United States
The Extracellular Oxygen Consumption kit is a tool used to measure the oxygen consumption rate of cells or tissue samples in an extracellular environment. The kit provides a standardized method for quantifying cellular respiration and metabolic activity.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using extracellular oxygen consumption kit
1
Extracellular Oxygen Consumption Assay
2
Mouse PH Fatty Acid Oxidation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse PHs were seeded and cultured in 96‐well plates overnight (1 × 104 cells per well). For the fatty acid oxidation assay, culture media were replaced by glucose‐deprived DMEM overnight. Oxygen consumption was measured by using an FAO kit (Abcam, Cambridge, MA, USA) and an Extracellular Oxygen Consumption kit (Abcam, Cambridge, MA, USA) according to the manufacturer's instructions.
3
Measuring Oxygen Consumption in Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Oxygen consumption of cell lines was measured using the fluorescence-based Extracellular Oxygen Consumption kit (Abcam) according to the manufacturer's protocol except for the NB-4 cell line. NB-4 cells were plated at a concentration of 3 × 10 6 cells/ml instead of 4 × 10 6 cells/ml. Cells were plated in duplicate and experiments performed three times for each cell line. Cells were treated with 5 μM CAPE. For positive and negative controls of oxygen consumption level, 1 μM electron transport chain (ETC) uncoupler agent FCCP (carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, (Abcam) and 1 μM oligomycin (ATP synthase inhibitor) (Abcam) (0.75 μM for NB-4) were used. FCCP and oligomycin concentrations were determined with a preliminary cell proliferation experiment. Fluorescence levels were measured on a molecular devices plate reader system for 2 h with an interval of 90 seconds at 37°C.
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!