Blood was collected from tail-vein using EDTA-coated tubes one-week post-irradiation. Cells were removed from plasma by centrifugation for 10 min at 2000× g at 4 °C. Supernatant was collected and stored at −80 °C. Plasma Growth Hormone (GH) titration was performed using a commercial ELISA kit (Rat/Mouse growth hormone Elisa kit EZRMGH-45K, Millipore-Sigma).
Ezrmgh 45k
Manufactured by Merck Group
Sourced in United States
The EZRMGH-45K is a laboratory equipment product manufactured by Merck Group. It is designed for general laboratory use. The core function of the EZRMGH-45K is to perform tasks related to sample processing and analysis. Detailed technical specifications and performance characteristics are not available.
Automatically generated - may contain errors
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16 protocols using ezrmgh 45k
1
Post-Irradiation Plasma GH Quantification
2
Quantifying Rat Growth Hormone in EV
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Rat GH concentration in culture medium and in isolated EV aliquots was assessed by enzyme-linked immunosorbent assay (ELISA) (Millipore Sigma, catalog No. EZRMGH-45K, RRID:AB_2892711, https://scicrunch.org/resolver/AB_2892711 ).
3
Quantification of Plasma Hormones
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Plasma insulin, IGF1/IGF2 and GH were determined by ELISA according to manufacturers’ instructions (10-1247-01, Mercodia; EMIGF1 and EMIGF2, Thermo Fisher; EZRMGH-45K, Millipore Sigma).
4
Mouse Growth and Hormone Profiles
5
Serum and Tissue GH-IGF1 Quantification
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Blood samples (n = 5 per genotype) were collected at the time of animal sacrifice, kept in ice for 30 min, and then centrifuged in an Eppendorf benchtop centrifuge for 15 min at 3,000 rpm to separate serum. Dissected liver and hippocampal tissues (n = 5 per genotype) were homogenized in lysis buffer (50 mM Tris–HCl pH 7.5, 1% NP-40, 1% Triton-100, 1 mM PMSF, 10% glycerol, and protease inhibitor cocktail; Sigma-Aldrich). Homogenates were incubated in ice for 30 min, centrifuged at 12,000 rpm for 5 min at 4°C, and supernatants were recovered and stored at −80°C. Protein concentration in serum, liver, and hippocampal samples was determined by BCA method (Pierce). Samples were processed for GH (EZRMGH-45K, Millipore) and IGF-1 (ELM-IGFI; RayBiotech) ELISA according to manufacturers’ protocol. All samples were analyzed in duplicate.
6
Measurement of Metabolic Biomarkers in Mice
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Blood glucose was assessed by glucometer (Accu‐Chek system; Roche Diagnostics, Barcelona, Spain). Gh (EZRMGH‐45K, sensitivity 0.07 ng/ml; Millipore, Billerica, MA, USA), insulin (EZRMI‐13K, sensitivity 0.2 ng/ml; Millipore), Igf1 (AC‐18F1, Immunodiagnostic Systems, sensitivity 63 ng/ml; Fountain Hills, AZ, USA), leptin (EZML‐82K, sensitivity 0.05 ng/ml; Millipore) and corticosterone (AC‐14F1, Sensitivity 0.55 ng/ml Immunodiagnostic Systems, Boldon, UK) levels were assessed using ELISA kits. Ten mice/group (for insulin determination) or 4–5 mice/group (for leptin, Gh, Igf1 and corticosterone determinations) were used. All details regarding the protocol, specificity, detectability and reproducibility for each assay can be accessed at the websites of the indicated companies.
7
IGF-1, GH, and Insulin Levels in Jazf1 Mice
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Blood samples were taken from the tail vein after 16 h overnight fast in Jazf1 KO and WT mice. Plasma IGF-1 and GH levels were measured using ELISA kits (22-IG1MS-E01, ALPCO, NH, USA and EZRMGH-45 K, Millipore, MA, USA), according to the manufacturer’s instructions. Plasma insulin levels were measured by radioimmunoassay (Linco). Plasma glucose measured by glucose analyzer (YSI).
8
Serum Hormone Profiling in Mice
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Serum/plasma were collected via orbital bleeding immediately after euthanasia with CO2 between 8 and 10 a.m. Hormones were measured by ELISA: GH (EZRMGH-45K; Millipore), IGF-1 (22-IG1MS-E01; ALPCO), leptin (EZRL-83K; Millipore), insulin (NC9440604; Mercodia), IGF-1 binding protein-3 (IGFBP-3; EMIGFBP3; Thermo Fisher Scientific), cytokines (K152A0H-1; Meso Scale Discovery), serum IGFBP-1 (CL0383; Cell Applications, Inc.), and IGFBP-2 (IRKTAH5375; Innovative Research, Inc.). Serum acid labile subunit (ALS) levels were determined by Western immunoblotting (AF1436; R&D Systems).
9
Fasting GH Levels in Oxr1A-/- Mice
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Circulating levels of GH were assayed by ELISA. We fasted 6-month-old WT and Oxr1A À/À mice, eight mice in each group, for 4 hours from 7 am. We collected blood samples at À30 and 5, 30, and 240 min from Lateral saphenous vein with Microvetteâ tubes (Microvetteâ CB 300 ml, K2 EDTA, violet US code, Sarstedt), 50 ml each time, and monitored glucose level before and after fasting. The blood samples were centrifuged at 12,000 rpm for 5 min at 4 C, and the plasma samples were collected and stored at À80 C. The plasma GH concentration was measured using commercially available ELISA kit (EZRMGH-45K, Millipore Co) according to the manufacturer's instructions. Absorbance for GH was measured and analyzed using a standard curve. We analyzed the individual provocative growth hormone response by calculation the ratio of peak and basal level of GH concentrations.
10
In vivo hormone measurement protocol
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For in vivo hormone measurements, blood from the subclavian vein was collected and centrifuged to obtain serum. GH, IGF-I and prolactin levels were measured by ELISA commercial kits (EZRMGH-45K, Merck Millipore, Madrid, Spain; AC-18F1, Immunodiagnostic Systems, Ortho Clinical Diagnostics, Rochester, USA; A05101, Bertin Pharma, Montigny le Bretonneux, France).
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