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Protocol for Isolation of Cardiomyocyte from Adult Mouse and Rat

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Fat Turnover Assay in Drosophila

Authors: Subhash D. Katewa and Pankaj Kapahi, date: 11/05/2016, view: 10638, Q&A: 0

[Abstract] Like all animals, Drosophila shows robust fat (triglyceride) turnover, i.e., they synthesize, store and utilize triglyceride for their daily metabolic needs. The protocol describes a simple assay to measure this turnover of triglycerides in Drosophila.

Spectrophotometric Determination of Glutamine Synthetase Activity in Cultured Cells

Authors: I-Chen Peng, Alex J. Bott and Wei-Xing Zong, date: 10/05/2016, view: 11761, Q&A: 0

[Abstract] Glutamine synthetase (GS), which catalyzes the conversion of glutamate and ammonia to glutamine, is widely distributed in animal tissues and cell culture lines. The importance of this enzyme is suggested by the fact that glutamine, the product of GS-catalyzed de novo synthesis reaction, is the most abundant free amino acid in blood (Smith and ...

Measurement of Transferrin- and Non-transferrin-bound Iron Uptake by Mouse Tissues

Authors: Supak Jenkitkasemwong, Chia-Yu Wang and Mitchell D. Knutson, date: 09/05/2016, view: 7614, Q&A: 0

[Abstract] Iron in blood plasma is bound to its transport protein transferrin, which delivers iron to most tissues. In iron overload and certain pathological conditions, the carrying capacity of transferrin can become exceeded, giving rise to non-transferrin-bound iron, which is taken up preferentially by the liver, kidney, pancreas, and heart. The ...

Quantification of 2-Hydroxyglutarate Enantiomers by Liquid Chromatography-mass Spectrometry

Authors: William M. Oldham and Joseph Loscalzo, date: 08/20/2016, view: 9734, Q&A: 0

[Abstract] Two enantiomers of 2-hydroxyglutarate (2HG), L (L2HG) and D (D2HG), are metabolites of unknown function in mammalian cells that were initially associated with separate and rare inborn errors of metabolism resulting in increased urinary excretion of 2HG linked to neurological deficits in children (Chalmers et al., 1980; Duran et al ...

Extraction and Quantification of Tryptophan and Kynurenine from Cultured Cells and Media Using a High Performance Liquid Chromatography (HPLC) System Equipped with an Ultra-sensitive Diode Array Detector

Authors: Jeffrey Kim, Benjamin Stewart and Robert H Weiss, date: 04/05/2016, view: 7521, Q&A: 0

[Abstract] Evidence of the involvement of tryptophan and its metabolite, kynurenine, in various biological processes including cancer is constantly expanding. Analysis of cell extracts and culture media can allow for quick snapshots of the metabolic fluctuations occurring in vitro. Here, we describe a method for metabolite extraction from mammalian ...

Measurement of 2-methylthio Modifications in Mitochondrial Transfer RNAs by Reverse-transcription Quantitative PCR

Authors: Fan-Yan Wei and Kazuhito Tomizawa, date: 01/05/2016, view: 7739, Q&A: 0

[Abstract] 2-Methylthio-N⁶-isopentenyladenosine (ms²i⁶A) is an evolutionally conserved posttranscriptional modification found at position 37 of four mammalian mitochondrial tRNAs, mt-tRNA^Ser(UCN), mt-tRNA^Trp, mt-tRNA^Phe and mt-tRNA^Tyr. The ms² modification in ms² ...

LDH-A Enzyme Assay

Authors: Di Zhao, Yue Xiong, Qun-Ying Lei and Kun-Liang Guan, date: 10/05/2013, view: 13736, Q&A: 0

[Abstract] LDH (Lactate dehydrogenase) enzyme catalyzes the reversible conversion of pyruvate to lactate using NAD+ as a cofactor. Although the physiological significance of lactate accumulation in tumor cells, a dead-end product in cellular metabolism, is currently a topic of debate, it has long been known that many tumor cells express a high level of LDH-A ...

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