Nori Rat Estrogen Receptor Alpha ELISA Kit Summary

Alternative names for estrogen receptor alpha: ERa, NR3A1, ESR1

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Estrogen receptor alpha (ERα), also known as NR3A1 (nuclear receptor subfamily 3, group A, member 1), is one of two main types of estrogen receptor that is activated by the sex hormone estrogen. ERα is encoded by the gene ESR1.^[1][2] The estrogen receptor (ER) is a ligand-activated transcription factor composed of several domains important for hormone binding, DNA binding, and activation of transcription. ERα is widely expressed throughout the body. The development and function of these tissues is disrupted in animal models lacking active ERα genes, such as the ERα knockout mouse (ERKO), providing a preliminary understanding of ERα function at specific target organs.^[3] ERα is essential in the maturation of the female reproductive phenotype. Activation of ERα is known to trigger cell proliferation in the uterus.^[4] The uterus of female ERKO mice is hypoplastic, suggesting that ERα mediates mitosis and differentiation in the uterus in response to estrogen stimulation.^[5] Similarly, prepubertal female ERKO mice develop ovaries that are nearly indistinguishable from those of their wildtype counterparts. However, as the ERKO mice mature they progressively present an abnormal ovarian phenotype in both physiology and function.^[5][3] Specifically, female ERKO mice develop enlarged ovaries containing hemorrhagic follicular cysts, which also lack the corpus luteum, and therefore do not ovulate.^[5][3] This adult ovarian phenotype suggests that in the absence of ERα, estrogen is no longer able to perform negative feedback on the hypothalamus, resulting in chronically elevated LH levels and constant ovarian stimulation.^[5] These results identify a pivotal role for ERα in the hypothalamus, in addition to its role in the estrogen-driven maturation through theca and interstitial cells of the ovary.^[5] Estrogen signaling through ERα appears to be responsible for various aspects of central nervous development, such as synaptogenesis and synaptic remodeling.^[3] Estrogen insensitivity syndrome is a very rare condition characterized by a defective ERα that is insensitive to estrogens.^[6] Genetic polymorphisms in the gene encoding the ERα have been associated with breast cancer in women, gynecomastia in men and dysmenorrhea.^[7]

References

1. Walter P, et al. (1985). Proc Natl Acad Sci USA. 82 (23): 7889–7893.
2. Greene GL, et al. (1986). Science. 231 (4742): 1150–1154.
3. Lee HR, et al. (2012). Laboratory Animal Research. 28 (2): 71–76.
4. Paterni I, et al. (2014). Steroids. 90: 13–29.
5. Curtis Hewitt S, et al. (2000). Breast Cancer Research. 2 (5): 345–352.
6. Smith EP, et al. (1994). The New England Journal of Medicine. 331 (16): 1056–1061.
7. Jahandoost S, et al. (2017) Journal of the National Medical Association. 109 (2): 126–138.