Serum insulin concentrations were measured using an ELISA kit from ALPCO Diagnostics (80‐INSMSU‐E01, Salem, NH) according to the manufacturer's instructions. Insulin resistance was assessed using the homeostasis model assessment of insulin resistance (HOMA‐IR). The HOMA‐IR was calculated as fasting insulin concentration (μU/mL) × fasting glucose concentration (mmol/L)/22.5. The serum cholesterol and triglyceride levels were measured using colorimetric quantitation kits (K603‐100 and K622‐100, Biovision Inc., Mountain View, CA), respectively. Each sample was measured in duplicate.
K603 100
Manufactured by Abcam
Sourced in United States
K603-100 is a laboratory equipment product. It is a 100 µl glass Hamilton syringe that can be used for accurate and precise liquid handling in various scientific applications.
Automatically generated - may contain errors
Lab products found in correlation
K622 100, by Abcam (6 mentions)
Mouse ultrasensitive insulin elisa kit, by ALPCO (2 mentions)
Au480, by Olympus (1 mentions)
Clarity bg1000, by Avantor (1 mentions)
47 adpms e01, by ALPCO (1 mentions)
Mmhmag 44k, by Merck Group (1 mentions)
Mf2100, by R&D Systems (1 mentions)
Tro100 1kt, by Merck Group (1 mentions)
K612 100, by Abcam (1 mentions)
E bc k235 m, by Elabscience (1 mentions)
E bc k236 m, by Elabscience (1 mentions)
Ezml 82k, by Merck Group (1 mentions)
Ab655390, by Abcam (1 mentions)
11 protocols using k603 100
1
Insulin Resistance and Lipid Profiles
2
Metabolic Profiling in Offspring Mice
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At 3 weeks of age, the offspring mice were euthanized, and the blood samples were collected. The blood samples were centrifuged at 4000× g for 10 min, and the serum was stored in aliquots at −80 °C. The serum insulin concentrations were measured using the Mouse Ultrasensitive Insulin ELISA kit from ALPCO Diagnostics (80-INSMSU-E01, Salem, NH, USA). The intra-assay coefficients of variation for the insulin measurements were 4.2%. The insulin sensitivity was assessed using the homeostasis model assessment of insulin resistance (HOMA-IR). The HOMA-IR was calculated as the fasting insulin concentration (μU/mL) × fasting glucose concentration (mmol/L)/22.5 [36 (link)]. The serum cholesterol (K603-100, kits from BioVision Inc., Mountain View, CA, USA) and triacylglycerol (K622-100, kits from BioVision Inc.) were measured by colorimetric methods.
3
Serum Lipid Analysis in Vmp1 Mice
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Six pairs of 18-month-old Vmp1flox/+;Villin-Cre mice and Vmp1flox/flox;Villin-Cre mice fed ad libitum were examined. Blood was allowed to stand for 2 hr and centrifuged at 1600 g for 15 min for collecting the serum. Serum samples were then measured by OLYMPUS AU480 automatic biochemical analyzer.
HepG2 cells were treated with siRNA twice as described above. HepG2 cells (approximately 1 × 105 cells) were cultured in serum-free medium for 24 hr before analysis. For total lipid extraction from culture medium, the Bligh and Dyer method was performed. Both extra- and intra-cellular cholesterol and triglyceride levels were measured using quantitation kits (K603-100 and K622-100, respectively, Biovision Inc) according to the manufacturer’s protocols.
HepG2 cells were treated with siRNA twice as described above. HepG2 cells (approximately 1 × 105 cells) were cultured in serum-free medium for 24 hr before analysis. For total lipid extraction from culture medium, the Bligh and Dyer method was performed. Both extra- and intra-cellular cholesterol and triglyceride levels were measured using quantitation kits (K603-100 and K622-100, respectively, Biovision Inc) according to the manufacturer’s protocols.
4
Comprehensive Metabolic Profiling Assay
5
Metabolic Assessment in Mice
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Serum insulin concentrations were determined using the Mouse Ultrasensitive Insulin ELISA kit from ALPCO Diagnostics (80-INSMSU-E01; Salem, NH, USA). Insulin sensitivity was assessed using the homeostasis model assessment of insulin resistance (HOMA-IR), which was calculated as the fasting insulin concentration (μU/mL) × fasting glucose concentration (mmol/L)/22.515 (link). The serum and hepatic lipid levels were measured using a colorimetric cholesterol quantitation kit (K603-100; Biovision Inc., Mountain View, CA, USA) and a colorimetric triacylglycerol quantitation kit (K622-100; Biovision Inc., Mountain View, CA, USA) according to the manufacturer's protocol16 (link),17 (link).
6
Cholesterol Quantification in AML12 Cells
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Total cholesterol concentration was measured using a cholesterol assay kit (K603-100; BioVision, Milpitas, CA). A total of 1 × 106 AML12 cells (untreated, or treated to reduce or enrich with cholesterol) were extracted with 200 μL chloroform:isopropanol:NP40 (7:11:0.1). After centrifugation for 10 minutes at 15,000 × g, the organic phase was transferred to a new tube, air-dried at 50°C to remove the chloroform, and vacuumed for 30 minutes to remove trace organic solvent. The dried lipids were dissolved with 200 μL cholesterol assay buffer and vortexed to homogeneity. A total of 1–50 μL of the extracted sample was used per assay and adjusted to a volume of 50 μL with cholesterol assay buffer. A total of 50 μL Reaction Mix containing 44 μL cholesterol assay buffer, 2 μL cholesterol probe, 2 μL cholesterol enzyme mix, and 2 μL cholesterol esterase were added to each sample for incubation at 37°C for 1 hour. Absorbance was measured at 570 nm using the Infinite 200 Spectrophotometer (Tecan Group AG).
7
Quantifying Cholesterol in CD8+ T Cells
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Cholesterol and esterified cholesterol levels in CD8+ T cells were evaluated using a cholesterol assay kit (K603‐100; BioVision). Briefly, cholesterol was extracted from human and mouse CD8+ T cells in accordance with the manufacturer's instructions. Then, changes in the cholesterol concentration after SCD1 inhibitor treatment were determined. A colorimetric assay was conducted at an optical density of 570 nm to determine the cholesterol content from a standard curve.
8
Quantification of Metabolic Markers
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Triglycerides, total cholesterol, and free fatty acids were quantified using commercial kits (triglycerides, K622-100; cholesterol, K603-100; free fatty acids, K612-100; Biovision, Milpitas, CA, USA). Serum insulin concentration was measured with an ELISA kit (#10-1247-01; Mercodia, Uppsala, Sweden), and another commercial kit (#32180; Immunodiagnostics Limited, Science Park, Hong Kong, China) was used in FGF21 measurement. Every sample was assayed in duplicate.
9
Liver Biochemical Marker Quantification
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The triglyceride and cholesterol contents in the liver were measured using a triglyceride quantification colorimetric kit (K622-100, Biovision, CA, United States) and a cholesterol quantification colorimetric kit (K603-100, Biovision, CA, United States) according to the manufacturer’s protocols. Serum alanine transaminase (ALT) and aspartate transaminase (AST) were detected using an ALT quantification kit (E-BC-K235-M, Elabscience, Wuhan, China) and an AST quantification kit (E-BC-K236-M, Elabscience, Wuhan, China) according to the manufacturer’s protocols.
10
Serum Biomarker Measurement Protocol
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After animals were anesthetized with pentobarbital sodium (50 mg/kg i.p.), whole blood samples were obtained from the vena cava. A whole blood sample was held for 30 min at room temperature to allow clotting. The sample was centrifuged at 2,000–3,000 g for 10 min at 4°C; the serum was transferred to separate tubes without disturbing blood clots and stored at −80°C until analysis. Commercial ELISA kits were used to measure serum levels of adiponectin (Millipore, EZMADP-60K), resistin (R&D, MRSN00), and leptin (Millipore, EZML-82K) (28 (link)). Similarly, total cholesterol levels were assessed using spectrophotometric assays (Biovision, K603-100) according to the manufacturer’s instructions (28 (link)).
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