Total testosterone levels were measured in plasma samples collected from each mouse using the testosterone parameter assay kit (R&D Systems, Minneapolis, MN, USA), per manufacturer’s instructions.
Testosterone parameter assay kit
Manufactured by R&D Systems
Sourced in United States
The Testosterone Parameter Assay Kit is a laboratory tool designed to quantify the levels of testosterone in biological samples. The kit utilizes an enzyme-linked immunosorbent assay (ELISA) technique to provide accurate and reliable measurements of testosterone concentrations.
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Lab products found in correlation
Automated glucometer, by Roche (1 mentions)
Multiskan mk3, by Thermo Fisher Scientific (1 mentions)
Estradiol parameter assay kit, by R&D Systems (1 mentions)
Estradiol elisa kit, by Cayman Chemical (1 mentions)
Progesterone elisa kit, by Cayman Chemical (1 mentions)
Elisa kit for follicle stimulating hormone, by Cloud-Clone (1 mentions)
Elisa kit for luteinizing hormone, by Cloud-Clone (1 mentions)
12 protocols using testosterone parameter assay kit
1
Mouse Plasma Testosterone Quantification
2
Fasting Glucose, Insulin, and Testosterone
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Blood samples were collected from the tail vein of the animals after they had fasted for 12 h, while they were still awake. Blood glucose levels during fasting were measured using an automated glucometer by Roche. Adhering to the supplied protocols, fasting insulin (FINS) levels were determined with a rodent insulin ELISA kit (Thermo Fisher Scientific, Catalog # ERINS), and testosterone concentrations were quantified using a Testosterone Parameter Assay Kit (R&D Systems, Catalog # KGE010).
3
Testosterone Levels in Castrated Rats
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Thirty-two male rats were randomly assigned to four groups: control group (38.3 mg/kg placebo microspheres, i.m.), castration group (rats were castrated and treated with 38.3 mg/kg placebo microspheres, i.m.), Zoladex® group (1.44 mg/kg goserelin, s.c.), and LY01005 group (1.44 mg/kg goserelin, i.m.), which would receive respective treatments once every 28 days for a total of three doses. Blood samples (approximate 200 μL) were collected on Day 0 (prior to dosing) and days 1, 4, 7, 14, 21, 28, 35, 42, 49, 56, 63, 70, 77 and 84 after injection. Blood samples without anticoagulation were centrifuged at 3,000 rpm for 15 min at room temperature. Serum testosterone levels were analyzed using the Testosterone Parameter Assay Kit (R&D Systems, Inc., United States) according to the manufacturer’s instructions.
4
Anthropometric and Endocrine Biomarker Assessment
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For each participant, we performed the following basic anthropometric measurements: weight, height, waist circumference, and hip circumference. Serum levels of adiponectin, leptin, resistin, tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β), IL-6, human monocyte chemoattractant protein-1 (MCP-1), and plasminogen activator inhibitor-1 (PAI-1) were measured by immunoenzymatic assays using HADCYMAG-61K and MILLIPLEX MAP Human Adipocyte Magnetic Bead Panel-Endocrine Multiplex Assay with a multi-factor detection instrument (MACPIX, LUMINEX). Serum total estradiol concentrations and serum FSH concentrations were measured using an enzyme-linked immunosorbent assay (DRG Estradiol ELISA, EIA-2693; ALPCO FSH ELISA, 11-FSHHU-E01). Total testosterone concentration was determined using a competitive enzyme immunoassay (R&D Testosterone Parameter Assay Kit, KGE010). The fully automatic multifunctional enzyme labeling instrument used was a MULTISKAN MK3 (Thermo Fisher Scientific, San Jose, CA, USA).
5
Murine OPG and RANKL Quantification
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Quantitative levels of murine OPG in the serum of p53+/+, p53+/−, and p53−/− C57BL/6 mice and in the conditioned medium of MSCs were determinate in triplicate by ELISA according to the manufacturer’s protocol (Mouse OPG Quantikine ELISA kit immunoassay, R&D). Quantitative levels of murine RANKL in serum of p53+/+, p53+/−, and p53−/− C57BL/6 were determined in triplicate by ELISA according to the manufacturer’s protocol (Mouse RANKL Quantikine ELISA kit immunoassay, R&D). Quantitative levels of human OPG in conditioned medium of hUC were determinate in triplicate by ELISA according to the manufacturer’s protocol (Human Osteoprotegerin TFRSF11b ELISA kit, Sigma-Aldrich). Quantitative levels of testosterone were determined following the protocol of the Testosterone Parameter Assay Kit (R&D system).
6
Quantification of Serum Testosterone and Estradiol in Mice
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The serum testosterone level of mice was determined using a competitive binding Testosterone Parameter Assay Kit (R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions with testosterone as the standard. The minimum detectable dose of testosterone for the ELISA kit was approximately 0.03 ng/mL. The following compounds were tested for their cross-reactivity (testosterone cross-reactivity was set as 100%): DHT (2.6%), AD (<0.1%), 17β-estradiol (<0.1%), and progesterone (<0.1%). On the other hand, the serum estradiol level of mice was determined using a competitive binding Estradiol Parameter Assay Kit (R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions with 17β-estradiol as the standard. The minimum detectable dose of estradiol was approximately 5 pg/mL. The following compounds were tested for cross-reactivity (17β-estradiol cross-reactivity was set as 100%): estrone (0.26%), estriol (0.86%), 17α-ethinylestradiol (<0.1%), and progesterone (<0.1%).
7
Testosterone Measurement in TM3 Cells
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Testosterone levels in the TM3 cell supernatant were measured using the Testosterone Parameter Assay Kit (KGE010 R&D Systems Inc., Minneapolis, MN, USA). Serum testosterone levels were measured using the Testosterone Parameter Assay Kit (MM 0410O2 MEIMIAN, Inc., Yancheng, China).
8
Serum Testosterone Regulation in Castrated Rats
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Forty-eight male rats were randomly assigned to six groups: control group (38.3 mg/kg placebo microspheres, i. m.), castration group (rats were castrated and treated with 38.3 mg/kg placebo microspheres, i.m.), Zoladex® group (1.44 mg/kg goserelin, s.c.), and LY01005 groups (0.36, 0.72 and 1.44 mg/kg goserelin, i. m.). Blood samples (approximate 200 μL) were collected on Day 0 (prior to dosing) and days 1, 2, 4, 7, 10, 14, 18, 21, 24, 28, 32 and 35 after injection. Blood samples without anticoagulation were centrifuged at 3,000 rpm for 15 min at room temperature. Serum testosterone levels were analyzed using the Testosterone Parameter Assay Kit (R&D Systems, Inc., United States) according to the manufacturer’s instructions.
9
Serum Testosterone Measurement by ELISA
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Serum testosterone was measured by the ELISA method with a testosterone parameter assay kit (R&D Systems, Minneapolis, MN, USA; cat. no. SKGE010) according to the manufacturer's protocol.
10
Testosterone Level in Rat Serum
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A total of 3 rats in every group were used to detect the hormone level in serum. The level of testosterone in rats was determined by ELISA, and the Testosterone Parameter Assay Kit (KGE010; R&D systems) was used to measure the testosterone level according to the manufacturer's instruction.
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