Difference between revisions of "Wensaas 2007 J Lipid Res"
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{{Publication | {{Publication | ||
|title=Wensaas AJ, Rustan AC, Lövstedt K, Kull B, Wikström S, Drevon CA, Hallén S (2007) Cell-based multiwell assays for the detection of substrate accumulation and oxidation. J Lipid Res 48:961-7. | |title=Wensaas AJ, Rustan AC, Lövstedt K, Kull B, Wikström S, Drevon CA, Hallén S (2007) Cell-based multiwell assays for the detection of substrate accumulation and oxidation. J Lipid Res 48:961-7. | ||
|info=[http://www.ncbi.nlm.nih.gov/pubmed/17213484 PMID: 17213484 Open Access] | |info=[http://www.ncbi.nlm.nih.gov/pubmed/17213484 PMID: 17213484 Open Access] | ||
|authors=Wensaas AJ, Rustan AC, Loevstedt K, Kull B, Wikstroem S, Drevon CA, Hallen S | |authors=Wensaas AJ, Rustan AC, Loevstedt K, Kull B, Wikstroem S, Drevon CA, Hallen S | ||
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|journal=J Lipid Res | |journal=J Lipid Res | ||
|abstract=We describe multiwell assays for detecting the accumulation as well as the subsequent oxidation of (14)C-labeled substrates in cultured cells. Accumulation is monitored in real time by an established scintillation proximity assay in which the scintillator is embedded in the plate base primarily detecting cell-associated radiolabel. The substrate oxidation assay is a novel variant of previously described experimental approaches aimed at trapping (14)CO(2) produced by isolated enzymes, organelles, or intact cells. This method uses a standard 96-well tissue culture plate and, on top, an inverted filter plate immersed with NaOH that are clamped into a sandwich sealed with a silicon gasket to obtain gas-tight compartments. (14)CO(2) is captured in the filter and quantified by conventional scintillation. We demonstrate both the accumulation and subsequent oxidation of (14)C-labeled substrates in cultured human myotubes, adipocytes, and hepatocytes. Both methods are adaptable for compound screening; at the same time, these protocols provide easy-to-use and time- saving methods for ''in vitro'' studies of cellular fuel handling. | |abstract=We describe multiwell assays for detecting the accumulation as well as the subsequent oxidation of (14)C-labeled substrates in cultured cells. Accumulation is monitored in real time by an established scintillation proximity assay in which the scintillator is embedded in the plate base primarily detecting cell-associated radiolabel. The substrate oxidation assay is a novel variant of previously described experimental approaches aimed at trapping (14)CO(2) produced by isolated enzymes, organelles, or intact cells. This method uses a standard 96-well tissue culture plate and, on top, an inverted filter plate immersed with NaOH that are clamped into a sandwich sealed with a silicon gasket to obtain gas-tight compartments. (14)CO(2) is captured in the filter and quantified by conventional scintillation. We demonstrate both the accumulation and subsequent oxidation of (14)C-labeled substrates in cultured human myotubes, adipocytes, and hepatocytes. Both methods are adaptable for compound screening; at the same time, these protocols provide easy-to-use and time- saving methods for ''in vitro'' studies of cellular fuel handling. | ||
|keywords=Fatty acids, Glucose uptake, CO2 capture, Scintillation proximity assay | |keywords=Fatty acids, Glucose uptake, CO2 capture, Scintillation proximity assay | ||
|mipnetlab=SE Molndal Hallen S | |||
}} | }} | ||
{{Labeling | {{Labeling | ||
Latest revision as of 16:18, 27 March 2018
| Wensaas AJ, Rustan AC, Lövstedt K, Kull B, Wikström S, Drevon CA, Hallén S (2007) Cell-based multiwell assays for the detection of substrate accumulation and oxidation. J Lipid Res 48:961-7. |
Wensaas AJ, Rustan AC, Loevstedt K, Kull B, Wikstroem S, Drevon CA, Hallen S (2007) J Lipid Res
Abstract: We describe multiwell assays for detecting the accumulation as well as the subsequent oxidation of (14)C-labeled substrates in cultured cells. Accumulation is monitored in real time by an established scintillation proximity assay in which the scintillator is embedded in the plate base primarily detecting cell-associated radiolabel. The substrate oxidation assay is a novel variant of previously described experimental approaches aimed at trapping (14)CO(2) produced by isolated enzymes, organelles, or intact cells. This method uses a standard 96-well tissue culture plate and, on top, an inverted filter plate immersed with NaOH that are clamped into a sandwich sealed with a silicon gasket to obtain gas-tight compartments. (14)CO(2) is captured in the filter and quantified by conventional scintillation. We demonstrate both the accumulation and subsequent oxidation of (14)C-labeled substrates in cultured human myotubes, adipocytes, and hepatocytes. Both methods are adaptable for compound screening; at the same time, these protocols provide easy-to-use and time- saving methods for in vitro studies of cellular fuel handling. • Keywords: Fatty acids, Glucose uptake, CO2 capture, Scintillation proximity assay
• O2k-Network Lab: SE Molndal Hallen S
Labels:
Organism: Human
Tissue;cell: Skeletal muscle, Liver, Fat
