Hepatic lipids were extracted using the Lipid Extraction kit (Cell Biolabs STA-612). Extracted lipids were air dried, resuspended in butanol, and triglycerides were quantified using a Triglyceride quantification kit (Cell Biolabs STA-396) according to the manufacture’s protocol. Hepatic glycogen quantification was measured using the Glycogen Assay Kit (Cell Biolabs STA-5022) according to the manufacture’s protocol.
Glycogen assay kit
Manufactured by Cell Biolabs
Sourced in United States
The Glycogen Assay Kit is a colorimetric assay designed to quantify glycogen levels in biological samples. The kit provides the necessary reagents to extract and hydrolyze glycogen, and then detect the resulting glucose using a coupled enzymatic reaction and colorimetric detection.
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Lab products found in correlation
Lipid extraction kit, by Cell Biolabs (1 mentions)
Triglyceride quantification kit, by Cell Biolabs (1 mentions)
Glucose assay kit, by Cell Biolabs (1 mentions)
Adenosine assay kit, by Cell Biolabs (1 mentions)
Total cholesterol assay kit, by Cell Biolabs (1 mentions)
Serum triglyceride quantification kit, by Cell Biolabs (1 mentions)
Free fatty acid assay kit, by Cell Biolabs (1 mentions)
6 protocols using glycogen assay kit
1
Hepatic Lipid and Glycogen Quantification
2
Glycogen Content Determination in C2C12 Cells
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To evaluate the effect of the samples on glycogen accumulation in C2C12 myoblasts, the glycogen content in the cells was determined using glycogen assay kit (Cell Biolabs, Inc., San Diego, CA, USA). Amyloglucosidase hydrolyzes glycogen to glucose and the glucose is oxidized by glucose oxidase, which produces hydrogen peroxide. The hydrogen peroxide is detected with a colorimetric probe. The color was measured at 540 nm using a microplate reader and glycogen content was calculated using a standard curve.
3
Insulin-Stimulated Glycogen Synthesis Assay
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Serum-starved H4IIEC3 cells were grown on gelatin coated culture dish (ϕ140 mm), and were pretreated with DMSO or 50 μM CK666 for 15 min and then stimulated by 1 μM insulin for 120 min in culture medium containing DMSO or 50 μM CK666, respectively. Next, the cells were trypsinized, resuspended, and then sonicated, followed by measurement of glycogen synthesis with a Glycogen Assay Kit (Fluorometric) (CELL BIOLABS, MET-5023). The mean and the standard error of the amount of glycogen are shown in the graph. Three independent experiments were performed.
4
Metabolic Response of Lepidopteran Larvae to Baculovirus Infection
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Third-instar larvae of S. litura and B. mori were individually injected with 1 × 106 PFU of AcMNPV or BmNPV. At 0, 24, and 48 hpi, the hemolymph and fat body were collected from the larvae. The concentration of trehalose, glycogen, glucose, adenosine in the hemolymph and fat body was determined by colorimetric methods using Trehalose Microplate Assay Kit (Cohesion Biosciences, Ltd., CAK1029), Glycogen Assay Kit (Cell Biolabs Inc., MET-5022), Glucose Assay Kit (Cell Biolabs Inc., STA-680), and Adenosine Assay Kit (Cell Biolabs Inc., MET-5090) respectively. Colorimetric methods using these commercially available kits have been established for insect samples in metabolic studies (Chang et al., 2020 (link); Lin et al., 2020 (link)). For each measurement, we added necessary positive and negative controls for color development according to the manufacturers' manuals to ensure the effectiveness of colorimetric assays. Uninfected larval samples were included as negative control groups in each experiment to weigh the accurate changes in metabolites in experimental groups.
5
Hepatic Glycogen Quantification
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For measurements of total glycogen in livers, we used the Glycogen Assay Kit (Cell Biolabs, Inc, San Diego, CA) following the manufacturer's protocol.
6
Hepatic Metabolite Profiling Protocol
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The hepatic metabolite levels for glycogen, triglyceride, non-esterified free fatty acid (NEFA) and total cholesterol were determined using commercially available kits from Cell Biolabs (CA, USA). Ultrasonic disruption in PBS containing 0.5% Triton X-100 was used to homogenize 200 mg of liver tissue, which was centrifuged at 5000 xg for 10 minutes at 4°C. The supernatant was collected, stored on ice, and used for glycogen and triglyceride quantification using the Glycogen Assay Kit (Cell Biolabs), and Serum Triglyceride Quantification Kit (Cell Biolabs), respectively. For cholesterol and free fatty acid quantification [26 (link)], 10 mg of liver tissue was extracted with 200 μL of a chloroform:isopropanol:NP-40 (7:11:0.1) mixture in a micro-homogenizer. The extract was centrifuged at 15,000 xg for 10 minutes, and the organic phase was transferred to a new tube and air dried at 50ºC to remove the chloroform. The dried lipids were dissolved and homogenized in 200 μL of 1X Assay Diluent via vortexing. Cholesterol and NEFA were quantified using Total Cholesterol Assay Kit (Cell Biolabs) and Free Fatty Acid Assay Kit (Cell Biolabs), respectively.
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