Species Reactivity
Mouse; Rat
Intended Use
The Estradiol ELISA is a competitive immunoassay for the quantitative measurement of estradiol in mouse/rat serum. For research use only.
Contents of Kit
1. SORB MT Microtiter Plate, 12 x 8 (break apart) strips with 96 wells; Wells coated with anti-Estradiol antibody.
2. CAL 0-5 Calibrators (Calibrator 0-5), 6 vials, lyophilized; Concentrations: 0, 5, 20, 80, 320, 1280 pg/mL. For reconstitution see "Reagent Preparation".
3. INC BUF Incubation Buffer, 1 vial, 7 mL, ready to use.
4. ENZ CONJ DIL Enzyme Conjugate Diluent, 1 vial, 30 mL, ready to use.
5. ENZ CONJ 100× Enzyme Conjugate, 1 vial, 0.3 mL (100× concentrated) 17β-Estradiol labeled horseradish peroxidase in buffered matrix. see "Reagent Preparation".
6. SUB TMB Substrate Solution, 1 vial, 22 mL, ready to use; contains tetramethylbenzidine (TMB) and hydrogen peroxide in a buffered matrix.
7. STOP SOLN Stop Solution, 1 vial, 7 mL, ready to use; contains 2 N hydrochloric acid solution.
8. WASH SOLN 10× Wash Solution, 1 vial, 50 mL (10× concentrated); see "Reagent Preparation".
Note: Additional Calibrator 0 for sample dilution is available upon request.
Storage
When stored at 2°C to 8°C unopened reagents will be stable until expiration date. Do not use reagents beyond this date. Opened reagents must be stored at 2°C to 8°C. After first opening, the reagents are stable for 30 days If used and stored properly. Microtiter wells must be stored at 2°C to 8°C. Take care that the foil bag is sealed tighty. Protect TMB-Substrate Solution from light Store Calibrators retrigerated, they will be stable at 2-8°C for 7 days after reconsttuton. For longer storage freeze at -20°C.
Precision
The intra-assay (within-run) variation was determined by 20 replicate measurements of three serum samples within one run.
The inter-assay (between-run) variation was determined by duplicate measurements of three serum samples in 11 different tests.
Detection Range
The range of the assay is between 2.5 – 1280 pg/mL.
Sensitivity
The lowest analytical detectable level of Estradiol that can be distinguished from the Zero Calibrator is 2.5 pg/mL at the 2SD confidence limit.
General Description
Estradiol (E2 or 17β-estradiol) is an estrogenic hormone produced by the ovaries and in smaller amounts by the adrenal cortex and testes. It is the most potent female sexual hormone and is essential for maintaining normal female functions. During the oestrous cycle which can be divided into four phases (proestrus, estrus, metestrus, diestrus) estradiol concentrations rise gradually from metestrus to proestrus and fall to barely detectable levels in estrus. The maximum concentration is reached around mid-day of proestrus (1,2). Apart from its effects on sexual characteristics it has important influence on the growth and development of the brain (3,4).
In female rodents, the determination of estradiol is a useful marker in evaluating and monitoring the state of the reproductive functions and pregnancy as well.
Standard Curve
Following data are intended for illustration only and should not be used to calculate results from another run.
Citations
Publication ()
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The immune and metabolic treatment approach of using testosterone on mice models of liver injury
Johnny Amer, Ahmad Salhab, Hadeel Snobar, Yazan Alhabil
Applications: ELISA
Reactive species: Mouse
"Abstract: Background: Natural killer (NK) cells showed an anti-fibrotic effect; however, their function is thought to be impaired in advanced liver injury. In the current study, we aimed to assess the immune and metabolic impact of testosterone on mice models of liver injury.
Methods: Carbon-tetrachloride induced liver fibrosis male mice models was i.p injected for 2 weeks (acute) and 4 weeks (chronic) (n = 36). Testosterone (4 mg/kg mouse body weight) was injected i.p. following the first week of the acute model of CCl4 and following the second week of the chronic model of CCl4. At the end of the experiments, mice were sacrificed, and serum was collected for assessing liver enzymes of ALT and AST, as well as inflammatory markers of IL-6, metabolic makers of C-peptide levels, and lipid and glucose profiles. Livers were harvested and used for histological assessments for inflammation and fibrosis. Fibrosis profiles from liver extracts, αSMA and Collagen III, were assessed by RT-PCR. Moreover, liver tissue-resident NK cells were isolated and evaluated for their activity by assessing INF-γ and IL-6 receptors using ELISA and flow cytometry, respectively."
Article snippet: Serum levels of testosterone and estradiol were assessed using * and Creative diagnostics: DEIA04927,respectively. 
Figure 1. Serum estradiol following testosterone treatment shown using ELISA.
COA
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