The β-oxidation rate of free fatty acid was measured as previously reported,63 (link) with some modifications. HepG2 cells were cultured in DMEM medium on 6-well plate to a density of 1×106 cells per well. For the assay, add to each well 1ml Krebs-Ringer phosphate buffer containing fatty acid-free BSA (20 mg·ml−1), glucose (2.5 mM), oleic acid (0.4 mM, Sigma), and 1-14C oleic acid (1.0 μCi·ml−1, GE Healthcare). To start the treatment, NEN (1.0 μM) or vehicle were added into each well, which was then covered by a piece of square filter paper soaked with hyamine hydroxide (Perkin Elmer). After incubation for 3 h, 0.5 ml of 4 M H2SO4 was added to each well and then the wells were sealed again for additional 30 min before the filter papers were carefully removed. The 14CO2 trapped on each filter paper was measured by a scintillation counter (Beckman LS 5000 TD Liquid Scintillation System).
Ls 5000 td liquid scintillation system
Manufactured by Beckman Coulter
The LS 5000 TD Liquid Scintillation System is a laboratory instrument used for the detection and measurement of low-level radioactivity in liquid samples. It employs the technique of liquid scintillation counting to quantify the presence of radioactive isotopes in a sample.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using ls 5000 td liquid scintillation system
1
Measuring Fatty Acid β-Oxidation in HepG2 Cells
2
Measuring Fatty Acid β-Oxidation in HepG2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The β-oxidation rate of free fatty acid was measured as previously reported,63 (link) with some modifications. HepG2 cells were cultured in DMEM medium on 6-well plate to a density of 1×106 cells per well. For the assay, add to each well 1ml Krebs-Ringer phosphate buffer containing fatty acid-free BSA (20 mg·ml−1), glucose (2.5 mM), oleic acid (0.4 mM, Sigma), and 1-14C oleic acid (1.0 μCi·ml−1, GE Healthcare). To start the treatment, NEN (1.0 μM) or vehicle were added into each well, which was then covered by a piece of square filter paper soaked with hyamine hydroxide (Perkin Elmer). After incubation for 3 h, 0.5 ml of 4 M H2SO4 was added to each well and then the wells were sealed again for additional 30 min before the filter papers were carefully removed. The 14CO2 trapped on each filter paper was measured by a scintillation counter (Beckman LS 5000 TD Liquid Scintillation System).
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!