Optic nerves were isolated from sacrificed mice and flash frozen in liquid N2. Hexokinase activity was quantified using a hexokinase assay kit (ScienCell), in accordance with the manufacturer’s instructions. Assay results were normalized to the total protein level in each ON sample as quantified with the Pierce BCA protein assay kit (ThermoFisher Scientific).
Hexokinase assay kit
Manufactured by ScienCell
Sourced in United States
The Hexokinase assay kit is a laboratory tool used to measure the activity of the enzyme hexokinase. Hexokinase plays a crucial role in glucose metabolism by catalyzing the phosphorylation of glucose to glucose-6-phosphate. This assay kit provides the necessary reagents and protocols to quantify hexokinase activity in biological samples.
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5 protocols using hexokinase assay kit
1
Quantifying Optic Nerve Hexokinase
2
Hexokinase Activity Assay with Compound AF
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HK activity was measured using a hexokinase assay kit (ScienCell Research Laboratories, Carlsbad, CA, USA) according to the assay procedures recommended by the manufacturer. Purified hexokinase was pre-incubated with various concentrations of AF for 10 min, and then mixed with the assay substrates to start the reaction. The changing NADPH absorbance was monitored in dark for 60 min, using a Multiskan plate reader (Thermo Scientific). For analysis of HK activity in cell lysates, A549 cells were treated with various concentrations of AF for 6 h, and protein extracts were prepared and immediately used for assay of HK activity using the substrate mixtures provided in the assay kit.
3
Hexokinase Activity Assay Protocol
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HK activity was measured using a hexokinase assay kit (ScienCell Research Laboratories, Carlsbad, CA, USA) according to the assay procedures recommended by the manufacturer. Briefly, cells were treated with AF (1 μmol/L), CE (10 μmol/L) or both for 24 h, and cell extracts were prepared and performed for assay of HK activity using the reagents provided in the assay kit.
4
Quantifying Cellular Hexokinase Activity
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HK2 activity was measured by using the ScienCell™ Hexokinase Assay Kit (Carlsbad, CA, USA). A total of 1×105 cells were collected and washed with PBS, and 100 μL of lysis buffer was added to each sample before incubation at 4 °C for 20 min. The samples were then centrifuged at 1000× g for 10 min to remove the insoluble material. Additionally, 10 μL of sample lysate was transferred to a 96-well microplate and mixed with 90 μL of Master Mix. The HK activity signal was measured immediately with a fluorescence plate reader and recorded for 20 min.
5
Quantifying Optic Nerve Hexokinase
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Optic nerves were isolated from sacrificed mice and flash frozen in liquid N2. Hexokinase activity was quantified using a hexokinase assay kit (ScienCell), in accordance with the manufacturer's instructions. Assay results were normalized to the total protein level in each ON sample as quantified with the Pierce BCA protein assay kit (ThermoFisher Scientific).
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