Livers of 4 h-fasted mice were assayed using the Hexokinase activity kit (MAK091, Sigma-Aldrich) according to the manufacturer's instructions.
Hexokinase activity kit
Manufactured by Merck Group
Sourced in United States
The Hexokinase Activity Kit is a laboratory tool designed to measure the enzymatic activity of hexokinase, a crucial enzyme involved in glucose metabolism. The kit provides the necessary reagents and protocols to quantify hexokinase activity in biological samples, enabling researchers to study glucose homeostasis and related metabolic processes.
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5 protocols using hexokinase activity kit
1
Hexokinase Activity in Fasted Mouse Livers
2
Hexokinase Activity Assay in Cells
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Cells were grown in six-well plates with media change every 24 h. When cells reached 80% confluency, hexokinase activity was assayed in cell homogenates using the Hexokinase activity kit (MAK091, Sigma, St. Louis, MO, USA) according to the manufacturer’s instructions. Activity was normalized to the amount of protein in the sample.
3
Hexokinase Activity Assay in MSCs
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Hexokinase activity was assayed in MSCs using the Hexokinase activity kit (MAK091, Sigma, St. Louis, MO, USA) according to the manufacturer's instructions. The MSCs were cultured in a 24-well plate. The cells were treated by TNFα and IFNγ for different time periods. MSCs were treated with inhibitors LY294002 (10 μM, MCE, USA) or triciribine (50 μM, MCE, USA) before addition of TNFα and IFNγ. The viruses of Myr-AKT and DN-AKT were added 72 h before addition of TNFα and IFNγ. The results were normalized to the amount of protein in the sample.
4
Hepatic Hexokinase Activity Assay
5
Quantifying Hexokinase Activity and Glycolysis
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To examine hexokinase activity a colorimetric assay was performed as per manufacturer's instructions (Hexokinase activity kit, Catalog #MAK091, Sigma). Sample preparation included lysis of 1×106 cells in assay buffer, with a 1:10 dilution of lysate used in assay. A glycolysis cell-based assay was performed to measure l-lactate production as per manufacturer's instructions (Catalog #600450, Caymen Chemical, Ann Arbor, MI, USA). 1×104 cells in 200 μL were seeded per well in a 96-well plate for 24 hours. 20 μL of medium was collected into a new 96 well plate for colorimetric detection. Absorbance at appropriate wavelengths was measured with a Synergy 2 microplate reader (BioTek, Winooski, VT, USA). Percent hexokinase activity and lactate production were determined by dividing the corrected absorbance reading for each replicate by the average corrected absorbance for the appropriate control.
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