Plasma samples were taken at the end of reperfusion period (at 120 min) just before intracardiac injection of Evans blue. The 17 β estradiol measurements were performed by using 17 beta Estradiol ELISA Kit (ab108667) of abcam (Cambridge, UK). The test was performed according to the manufacturer’s instructions using plasma samples of mice after I/R.
17 beta estradiol elisa kit
Manufactured by Abcam
Sourced in United Kingdom
The 17 beta Estradiol ELISA kit is a quantitative in vitro diagnostic test used to measure the concentration of 17 beta-estradiol in biological samples, such as serum or plasma. It employs a competitive enzyme immunoassay technique to determine the level of 17 beta-estradiol.
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Lab products found in correlation
8 protocols using 17 beta estradiol elisa kit
1
Plasma 17β-estradiol Measurement after I/R
2
Quantification of Plasma Estradiol and IL-1β
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The mouse blood was collected via a cardiac puncture in microtainer K2E (BD, Cat # 365974, Franklin Lakes, NJ, USA) and centrifuged at 1600× g for 15 min at 4 °C to measure the plasma 17β-estradiol levels. The supernatant plasma was collected, and the levels of 17β-estradiol were assessed using the 17 beta Estradiol ELISA kit (Abcam, Cat # ab108667, Cambridge, UK) following the manufacturer’s protocol. To measure the IL-1β release from the GT1b-stimulated primary mixed glia, we treated ATP 30 min prior to supernatants collection to induce the secretion of IL-1β, and collected supernatants of mixed glia centrifuged at 500× g for 5 min to discard the cell debris. The IL-1β levels were measured using the mouse IL-1 beta Quantikine ELISA kit (R&D Systems Inc., Cat # MLB00C, Minneapolis, MN, USA) following the manufacturer’s protocol.
3
Plasma Glucose, Insulin, and Estrogen Levels in Tumor-Bearing Animals
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Plasma glucose and insulin levels were measured from samples collected at sacrifice from non-fasting tumor-bearing animals, as previously described [5 (link)]. Plasma glucose levels were determined by OneTouch UltraMini (LifeScan, Inc., Milpitas, CA, USA) and the insulin levels were determined with the rat/mouse insulin ELISA kit (EMD Millipore), according to the manufacturer’s instructions. Plasma estrogen was measured with the 17-beta-estradiol ELISA Kit (Abcam).
4
Quantification of Serum Hormones
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Serum estradiol concentrations were quantified using a 17 beta Estradiol ELISA kit (ab 108667, Abcam, Cambridge, United Kingdom) according to the instructions. The minimum detected concentration of the kit was 8.68 pg/ml.
Serum leptin concentrations were quantified using a Leptin mouse ELISA kit (ab 100718, Abcam, Cambridge, United Kingdom) according to the given instructions. The minimum detected concentration of the kit was 4 pg/ml for leptin.
Serum DCA concentrations were quantified using an ELISA kit (CESO89Ge, Cloud-clone, China) according to the given instructions. The minimum detected concentration of the kit was 4.99 ng/ml for DCA.
Serum leptin concentrations were quantified using a Leptin mouse ELISA kit (ab 100718, Abcam, Cambridge, United Kingdom) according to the given instructions. The minimum detected concentration of the kit was 4 pg/ml for leptin.
Serum DCA concentrations were quantified using an ELISA kit (CESO89Ge, Cloud-clone, China) according to the given instructions. The minimum detected concentration of the kit was 4.99 ng/ml for DCA.
5
Estradiol Quantification from Serum
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Under deep anesthesia, blood was obtained from the inferior vena cava. Coagulated blood was centrifuged at 13,000× g for 10 min to separate serum. Estradiol concentration was determined with the 17-beta Estradiol ELISA Kit (ab108667, Abcam, Cambridge, UK) following the manufacturer’s manual. The optical density of the samples was measured by VICTOR X3 (Perkin-Elmer Korea, Seoul, Korea) microplate reader.
6
Serum Estradiol Quantification using ELISA
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Circulating estrogen levels were quantified according to the manufacturer’s protocol using the 17 beta Estradiol ELISA Kit (cat no. ab108667, Abcam). This kit was validated by Marino FE et al.80 (link). Briefly, 25 µl of blood serum samples, prepared standards, or controls were added in duplicate to a 96 well plate. 200 µl of the 17 beta Estradiol-HRP Conjugate were added to each well followed by incubation at 37 °C for two hours. Samples, standards, and controls were aspirated, and wells were washed three times with 300 µl of diluted washing solution (soak time >5 seconds) using an automated plate washer (BioTek 50TS). After washing, 100 µl tetramethylbenzidine (TMB) Substrate Solution was added to each well, and the plate was incubated for 30 minutes in the dark at room temperature. After incubation, 100 µl of Stop Solution was added into all wells in the same order and at the same rate as the substrate solution. Absorbance was measured at 450 nm with Spectramax M3 plate reader (Molecular Devices) within 30 minutes of adding the Stop Solution. Construction of standard curve and subsequent analyses were performed in Microsoft Excel.
7
Serum hormone quantification in mice
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Serum estradiol concentrations were quantified using an 17 beta Estradiol ELISA kit (ab 108664, abcam) according to the instructions. The minimum detected concentration of the kit was 8.68 pg/ml for EST.
Serum leptin concentrations were quantified using an Leptin mouse Elisa kit (ab 100718, Abcam) according to the instructions. The minimum detected concentration of the kit was 4 pg/mL for leptin.
Serum luteinizing hormone (LH) concentrations and follicle stimulating hormone (FSH) concentrations were quantified using LH mouse Elisa kit (AB-C4424B, Abmart) and FSH mouse Elisa kit (AB-3291A, Abmart) according to the instructions.
Serum leptin concentrations were quantified using an Leptin mouse Elisa kit (ab 100718, Abcam) according to the instructions. The minimum detected concentration of the kit was 4 pg/mL for leptin.
Serum luteinizing hormone (LH) concentrations and follicle stimulating hormone (FSH) concentrations were quantified using LH mouse Elisa kit (AB-C4424B, Abmart) and FSH mouse Elisa kit (AB-3291A, Abmart) according to the instructions.
8
Quantification of Steroid Hormones
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During the culture period at days 2, 4 and 6, spent media were collected and stored at -80°C for subsequent 17 beta-estradiol and progesterone quantification. 17 betaestradiol and progesterone levels were evaluated using 17 beta-estradiol ELISA kit (Abcam) and progesterone ELISA kit (Abcam), respectively, according to the manufacturer's instructions. Absorbance was calculated at 450 nm. The analytical sensitivity of the assay was 8.68 pg/mL (assay range, 20-2000 pg/mL) for 17 beta-estradiol and 0.05 ng/mL (assay range, 0.2-40 ng/mL) for progesterone.
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