Rat Estradiol ELISA Assay


The Eagle Biosciences Rat Estradiol ELISA Assay Kit is intended for the quantification of Rat Estradiol in serum. The Eagle Biosciences Rat Estradiol ELISA Assay Kit is for research use only and should not be used for diagnostic procedures.

Rat Estradiol ELISA Assay

The Rat Estradiol ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 2.5 pg/mL
Dynamic Range: 5-1280 pg/mL
Incubation Time: 2.5 hours
Sample Type: Rat Serum
Sample Size: 75 µL
Alternative Names Rat E2 ELISA

Assay Background

Estradiol (E2), also spelled oestradiol, is a steroid, an estrogen, and the primary female sex hormone. It is named for and is important in the regulation of the estrous and menstrual female reproductive cycles. Estradiol is essential for the development and maintenance of female reproductive tissues such as the breasts, uterus, and vagina during puberty, adulthood, and pregnancy, but it also has important effects in many other tissues, including bone, fat, skin, liver, and the brain. While estrogen levels in men are lower compared to those in women, estrogens have essential functions in men, as well. It is found in most vertebrates and crustaceans, insects, fish, and other animal species. Estradiol is produced especially within the follicles of the female ovaries, but also in other endocrine (i.e., hormone-producing) and nonendocrine tissues (e.g., including fat, liver, adrenal, breast, and neural tissues). Estradiol is biosynthesized from cholesterol through a series of chemical intermediates. One principal pathway involves the generation of androstenedione, which is converted into estrone by aromatase and then by 17β-hydroxysteroid dehydrogenase into estradiol. Alternatively, androstenedione can be converted into testosterone, an androgen and the primary male sex hormone, which in turn can be aromatized into estradiol. 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.

Related Products

Estradiol ELISA Assay Kit
Total Estrogens ELISA Assay

Additional Information

Assay Principle

This assay employs the competitive enzyme immunoassay technique. An antibody specific for Estradiol has been pre-coated onto a microtiter plate. Endogenous Estradiol in samples or standards competes with an Estradiol-horseradish peroxidase conjugate for binding to the coated antibody. After incubation the unbound conjugate is washed off. The amount of bound peroxidase conjugate is inversely proportional to the concentration of Estradiol in the sample. After addition of the substrate solution, the intensity of color developed is inversely proportional to the concentration of Estradiol in the sample. Estradiol concentration in the sample is calculated through a calibration curve.

Assay Procedure

  1. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal it.
  2. Add 75 μl of standards and samples in duplicate into appropriate wells.
  3. Add 50 μl of Incubation Buffer into each well, mix well. Incubate for 2 hours at room temperature on a microplate mixer or shaker (600rpm).
    Immediately before use dilute the HRP-conjugated Estradiol 1:100 in HRP- conjugated Estradiol Diluent.
  4. Add 50 μl of 1X HRP-conjugated Estradiol into each well, mix well.
  5. Incubate for 1 hour at room temperature on a microplate mixer or shaker (600rpm).
  6. Aspirate each well and wash, repeating the process 3 times for a total 4 washes. Wash by filling each well with 1× Wash Buffer (300 μl) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating, decanting or blotting against clean paper towels.
  7. Add 200 μl of TMB substrate to each well. Incubate for 30 minutes at room temperature in dark.
  8. Add 50 μl of Stop Solution to each well.
  9. Read the OD with a microplate reader at 450 nm immediately. It is recommended to read the wells within 15 minutes

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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