Mice were fasted for 16 h prior to glucose measurement and plasma collection. Glucose concentrations were measured using glucose strips and a glucometer (Lifescan One Touch Ultra). Insulin levels were measured using a mouse ultrasensitive insulin ELISA kit (Crystal Chem, 90080). Serum triglycerides of fasted mice and liver triglycerides from fed mice were measured using a triglyceride assay kit (Cayman Chemical, 10010303). GTT was performed in 16-h-fasted mice using 1.5 g/kg glucose interperitoneal injection. Serum alanine aminotransferase measurements were performed by IDEXX BioResearch Laboratory.
Mouse ultrasensitive insulin elisa kit
Manufactured by Crystal Chem
Sourced in United States
The mouse ultrasensitive insulin ELISA kit is a laboratory tool used to detect and measure insulin levels in mouse samples. It employs the enzyme-linked immunosorbent assay (ELISA) technique to provide highly sensitive and quantitative results.
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Lab products found in correlation
Ascensia breeze glucometer, by Bayer (2 mentions)
Nefa kit, by Fujifilm (2 mentions)
Infinity triglycerides kit, by Thermo Fisher Scientific (2 mentions)
Aspartate aminotransferase activity kit, by Abcam (2 mentions)
Alanine aminotransferase activity kit, by Abcam (2 mentions)
Mouse leptin elisa, by Crystal Chem (2 mentions)
Triglyceride assay kit, by Cayman Chemical (1 mentions)
Onetouch ultra, by LifeScan (1 mentions)
Qubit protein assay kit, by Thermo Fisher Scientific (1 mentions)
Prism v9, by GraphPad (1 mentions)
Triglyceride quantification colorimetric fluorometric kit, by Merck Group (1 mentions)
Mouse adiponectin elisa kit, by Crystal Chem (1 mentions)
Tac colorimetric assay kit, by Abcam (1 mentions)
Ethanol assay kit, by Abcam (1 mentions)
Cobas 8000, by Roche (1 mentions)
Mouse ultrasensitive insulin elisa kit, by ALPCO (1 mentions)
12 protocols using mouse ultrasensitive insulin elisa kit
1
Metabolic Profiling in Fasted Mice
2
Glucose Tolerance Test for Metabolic Evaluation
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For GTT, animals fasted for eight hours and then given an intraperitoneal injection of 2 g glucose per kg body weight. Blood glucose was measured by Ascensia Breeze Glucometer (Bayer) at 0, 20, 30, 60, 90, 120, 150, 180 and 210 min. intervals via tail vein puncture method. In our experiment, the GTT was performed at two-time points; at the age of 12–14 weeks (pre GC7 administration) and another at the age of 25–26 weeks (post GC7 administration). Following the glucose challenge, we also measured plasma insulin concentrations at 0, 10 and 30 min. using a mouse ultrasensitive insulin ELISA kit (Crystal Chem, Inc., USA).
3
Glucose Tolerance and Insulin Secretion Assay
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Fasting blood glucoses were measured weekly by tail vein nicking. Mice were also subjected to the glucose tolerance test (GTT) where animals fasted for 8–10 h before being administered an intraperitoneal injection of glucose (2 g/kg body weight). Blood glucose concentrations were measured at 0, 20, 30, 60, 90, 120, 150, 180, and 210 min using the tail vein nicking technique, and blood glucose was measured with an Ascensia Breeze Glucometer (Bayer). Simultaneously, at glucose challenge, serum insulin concentrations (GSIS) were measured at 0, 2, 10, and 30 min. The insulin concentration was measured by a mouse ultrasensitive insulin ELISA kit (Crystal Chem, Inc.) (Imam et al., 2019 (link), 2020 , 2021 (link); Imam and Jaume, 2020 ).
4
Plasma Metabolic Biomarker Analysis
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Whole blood was taken from the facial vein and blood glucose was measured with a glucose meter (Easy Touch) from the tail vein. Plasma was collected after centrifugation at 1,200 x rpm, 4°C for 10 min. Plasma triglyceride (TG) and free fatty acid (FFA) levels were measured with Infinity™ Triglycerides kit (Thermo Fisher) and NEFA kit (WAKO). Plasma insulin levels were measured with the Mouse Ultrasensitive Insulin ELISA kit (Crystal Chem, #90080), and leptin levels were measured with Mouse Leptin ELISA (Crystal Chem, #90030) kit. Plasma AST and ALT activity was measured with the Aspartate Aminotransferase Activity kit (Biovision, #K753) and Alanine Aminotransferase Activity kit (Biovision, #K752).
5
Insulin Extraction and Quantification
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Insulin was extracted from the pancreas (PN) by a standard acid-ethanol extraction method as described earlier51 (link). Insulin concentration in the extract was determined using a Mouse Ultrasensitive Insulin ELISA kit (Crystal Chem, Inc., USA). The insulin content was standardized to the total protein concentration measured by Qubit Protein Assay kit (Invitrogen).
6
Insulin Dynamics During Glucose Tolerance Test
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Blood samples were collected from the tail vein at 0 minutes and 15 minutes during the oGTTs after the glucose load. Serum insulin levels were subsequently determined using the mouse Ultra Sensitive Insulin ELISA Kit (Crystal Chem, Chicago, IL, USA).
7
Comprehensive Metabolic Profiling of Mice
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Whole blood was taken from the facial vein and blood glucose was measured with a glucose meter (Easy Touch) from the tail vein. Plasma was collected after centrifugation at 1,200g at 4 °C for 10 min. Plasma TG and FFA levels were measured with an Infinity Triglycerides kit (Thermo Fisher) and NEFA kit (WAKO). Plasma insulin levels were measured with the Mouse Ultrasensitive Insulin ELISA kit (Crystal Chem, 90080) and leptin levels were measured with a Mouse Leptin ELISA (Crystal Chem, 90030) kit. Plasma AST and ALT activity was measured with the Aspartate Aminotransferase Activity kit (Biovision, K753) and Alanine Aminotransferase Activity kit (Biovision, K752).
8
Quantifying Metabolic Biomarkers in ob/ob Mice
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Fasting insulin and triglycerides from blood collected at euthanasia were quantified using a mouse ultrasensitive insulin ELISA kit (Crystal Chem, Elk Grove Village, IL) and a Triglyceride Quantification Colorimetric/Fluorometric Kit (Sigma).
2.9 Histology. Histological analysis of pancreata was conducted per established methods, methodological details can be found in Supplementary Information.
2.10 Genotyping. Formalin fixed liver tissue was used to genotype each animal to confirm the presence or absence of the ob/ob mutation. Samples were sent to Transnetyx (Cordova, TN) and results can be found in Supplementary Information.
2.11 Statistics. Data were analyzed using GraphPad Prism v9.1.2 (La Jolla, CA). When values were missing, a mixed-effects model was used. When initial one-way or two-way ANOVA indicated a significant overall treatment effect or interaction (P<0.05), individual means were compared using Tukey's or Sidak's post-hoc tests to control for multiple comparisons (α = 0.05).
2.9 Histology. Histological analysis of pancreata was conducted per established methods, methodological details can be found in Supplementary Information.
2.10 Genotyping. Formalin fixed liver tissue was used to genotype each animal to confirm the presence or absence of the ob/ob mutation. Samples were sent to Transnetyx (Cordova, TN) and results can be found in Supplementary Information.
2.11 Statistics. Data were analyzed using GraphPad Prism v9.1.2 (La Jolla, CA). When values were missing, a mixed-effects model was used. When initial one-way or two-way ANOVA indicated a significant overall treatment effect or interaction (P<0.05), individual means were compared using Tukey's or Sidak's post-hoc tests to control for multiple comparisons (α = 0.05).
9
Glucose Tolerance Test in Mice
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Fasted (17 h) mice were intraperitoneally injected with D-glucose at a dose of 2.5 g/kg body weight. Whole venous blood was obtained from the tail at the indicated time points after glucose load and serum insulin was measured by Ultrasensitive Insulin Mouse Elisa Kit (CrystalChem).
10
Measuring Non-Fasting Glucose and Insulin
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Non-fasting blood glucose was measured in samples of cardiac blood by an Accu-Chek Diabetes kit (Roche Diagnostics, Indianapolis, IN, USA). Insulin levels (non-fasting) were measured in plasma samples using an ultra-sensitive insulin mouse ELISA kit (Crystal Chem Inc., IL, USA) according to the manufacture’s protocol.
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