Levels of insulin and C-peptide were determined in blood samples using the Rat/Mouse Insulin (Cat. #EZRMI-13K, Merck Millipore) and the mouse C-peptide (Cat. #80-CPTMS-E01, Alpco) ELISA kits, respectively.
Rat mouse insulin
Manufactured by Merck Group
Sourced in United States
Rat/Mouse Insulin is a laboratory reagent used for the measurement of insulin levels in rat and mouse samples. It is a highly specific and sensitive tool for researchers to quantify insulin concentrations in biological samples, such as plasma or tissue extracts, from rodent models. The product provides a reliable and accurate method to support research in areas related to insulin metabolism, diabetes, and other endocrine disorders.
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Lab products found in correlation
80 cptms e01, by ALPCO (1 mentions)
Accu check aviva plus, by Roche (1 mentions)
Ssofast evagreen supermix, by Bio-Rad (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Iscript reverse transcription supermix, by Bio-Rad (1 mentions)
Adiponectin, by Merck Group (1 mentions)
Accu chek, by Roche (1 mentions)
Leptin elisa kit, by R&D Systems (1 mentions)
Accu chek aviva, by Roche (1 mentions)
Accu check, by Roche (1 mentions)
7 protocols using rat mouse insulin
1
Insulin and C-Peptide Quantification
2
Measuring Glucose, Insulin, and Leptin
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Blood glucose levels were measured using an AccuCheck Aviva-Plus glucometer (Roche Diagnostics, Indianapolis, IN, USA) [24 (link)]. Serum insulin and leptin concentrations were analyzed using EMD Millipore Rat/Mouse Insulin (prod. no. EZRMI-13K) and Rat Leptin (prod. no. EZRL-83K) enzyme-linked immunosorbent assay kit reagents. The insulin assay was characterized by sensitivity of 0.1 ng/mL, with intra- and inter-assay coefficients of variation (CV) 3.22% and 6.95%, respectively. Leptin assay sensitivity was 0.08 ng/mL; intra- and extra-assay CVs were 2.49% and 3.93%.
3
Pioglitazone Effects on Metabolic Markers
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Pioglitazone was purchased from Takeda Pharmaceutical (Osaka, Japan). Rat/mouse insulin, adiponectin and leptin ELISA Kits were obtained from Millipore (Life Sciences/Biotech, Petaluma, MA, USA). Trizol® reagent was obtained from Invitrogen (Life Techologies, Eugene, OR, USA). iScript Reverse Transcription Supermix and Ssofast EvaGreen Supermix were obtained from BIO-RAD (Hercules, CA, USA). The hand-held glucometer (ACCU-CHEK®) was purchased from Roche diagnostics (Mannheim, Germany).
4
Serum Metabolic Biomarker Profiling
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Blood samples were obtained quickly after sacrifice by cardiac puncture, collected in tubes and centrifuged at 3000 r/min for 10 min. at 4°C. Serum was collected and stored at −80°C. Glucose was determined using a colorimetric commercial kit (Sigma-Aldrich).
Plasma insulin concentrations were measured in duplicate by a rat/mouse insulin enzyme-linked immunosorbent assay (ELISA) kit, using rat insulin as the standard (Millipore Co.Vimodrone, Italy) according to the manufacturer’s instructions. The intra-assay coefficient of variability (CV) was 1.17–3.22%, the inter-assay CV was 6.71–9.23%, and the sensitivity limit was 0.1 ng/mL.
Leptin concentrations were measured using a leptin ELISA kit (R&D Systems, UK) according to the manufacturer’s instructions. The intra-assay CV was 3.8–4.3%, the inter-assay CV was 5–7.6%, and the sensitivity limit was 22 pg/mL. Quantification of free fatty acids (FFA) was performed using a commercially available ELISA kit (Bioassay Tech. Lab., China) according to the manufacturer’s instructions. The intra-assay CV was <10%, inter-assay CV was <12%, sensitivity was 2.51 mM/L.
Plasma insulin concentrations were measured in duplicate by a rat/mouse insulin enzyme-linked immunosorbent assay (ELISA) kit, using rat insulin as the standard (Millipore Co.Vimodrone, Italy) according to the manufacturer’s instructions. The intra-assay coefficient of variability (CV) was 1.17–3.22%, the inter-assay CV was 6.71–9.23%, and the sensitivity limit was 0.1 ng/mL.
Leptin concentrations were measured using a leptin ELISA kit (R&D Systems, UK) according to the manufacturer’s instructions. The intra-assay CV was 3.8–4.3%, the inter-assay CV was 5–7.6%, and the sensitivity limit was 22 pg/mL. Quantification of free fatty acids (FFA) was performed using a commercially available ELISA kit (Bioassay Tech. Lab., China) according to the manufacturer’s instructions. The intra-assay CV was <10%, inter-assay CV was <12%, sensitivity was 2.51 mM/L.
5
Fasted Mice Insulin and Glucose
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Both plasma insulin and blood glucose were measured in mice that were fasted for six hours. Insulin concentration was measured using an ELISA insulin assay (Rat/Mouse Insulin Millipore). Plasma glucose concentration was determined with the use of an AccuChek Aviva glucometer (Roche, Germany).
6
Blood Glucose and Insulin Sensitivity Assay
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Blood glucose concentration was measured using the AccuChek glucometer. The plasma insulin concentration was determined by means of an ELISA insulin assay (Rat/Mouse Insulin, Millipore, Burlington, MA, USA). The insulin sensitivity was assessed using the homeostasis model assessment of insulin resistance (HOMA)-IR index using the formula [18 (link)]:
7
OGTT, ITT, and HOMA-IR Assessment in Rats
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OGTT was performed in by oral glucose administration at a dose of 3 g/kg. ITT was performed by intraperitoneal injection of insulin in a dose of 0.75 U/kg body weight. Blood samples from tail vein were measured at given intervals by glucometer (AccuCheck, Roche. Germany). Plasma insulin was measured with an ELISA insulin assay (Rat/Mouse Insulin Millipore). HOMA-IR index value was calculated according to Cacho et al. [24] .
Principal component analysis, correlation analysis and statistical significance estimation PCA was performed using Statistica 10.0 software package as described earlier [25] . To prevent an artificial increase in the PCA model strength, we excluded a majority of closely interdependent variables. We used Pearson's approach with Bonferroni correction for multiple comparisons to establish relationships between selected variables chosen on the basis of PCA analysis. Statistical significance between groups was estimated using ANOVA with the Tukey honestly significant difference post-hoc test for unequal n-numbers. Significance level was set to p < 0.05.
Principal component analysis, correlation analysis and statistical significance estimation PCA was performed using Statistica 10.0 software package as described earlier [25] . To prevent an artificial increase in the PCA model strength, we excluded a majority of closely interdependent variables. We used Pearson's approach with Bonferroni correction for multiple comparisons to establish relationships between selected variables chosen on the basis of PCA analysis. Statistical significance between groups was estimated using ANOVA with the Tukey honestly significant difference post-hoc test for unequal n-numbers. Significance level was set to p < 0.05.
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