ELISA Kit for Estrogen Receptor Alpha (ERa) Rattus norvegicus (Rat) Sandwich ELISA

ER-A1; ER; ESR; ESR1; ESRA; NR3A1; NR3-A1; Estrogen Receptor 1; Nuclear Receptor Subfamily 3,Group A,Member 1

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  • ELISA Kit for Estrogen Receptor Alpha (ERa) Packages (Simulation)
  • ELISA Kit for Estrogen Receptor Alpha (ERa) Packages (Simulation)
  • ELISA Kit for Estrogen Receptor Alpha (ERa) Results demonstration
  • SEB050Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Estrogen Receptor Alpha (ERa) and the recovery rates were calculated by comparing the measured value to the expected amount of Estrogen Receptor Alpha (ERa) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-104 101
EDTA plasma(n=5) 94-104 101
heparin plasma(n=5) 82-95 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Estrogen Receptor Alpha (ERa) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Estrogen Receptor Alpha (ERa) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Estrogen Receptor Alpha (ERa) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 93-102% 93-105% 80-103% 79-97%
EDTA plasma(n=5) 88-95% 88-101% 93-101% 81-97%
heparin plasma(n=5) 83-102% 79-89% 78-99% 80-95%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Estrogen Receptor Alpha (ERa)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Estrogen Receptor Alpha (ERa). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Estrogen Receptor Alpha (ERa). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Estrogen Receptor Alpha (ERa), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Estrogen Receptor Alpha (ERa) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Neuroprotective action of raloxifene against hypoxia-induced damage in mouse hippocampal cells depends on ERα but not ERβ or GPR30 signallingPubmed:24846829
  • Differential expression of estrogen receptor α and β isoforms in multiple and solitary leiomyomasPubMed: 26529545
  • Bisphenol A exposure alters release of immune and developmental modulators and expression of estrogen receptors in human fetal lung fibroblastsscience:S1001074216300924
  • Influenza A virus NS1 protein-induced JNK activation and apoptosis are not functionally linked.pubmed:28076660
  • Depressive-like effect of prenatal exposure to DDT involves global DNA hypomethylation and impairment of GPER1/ESR1 protein levels but not ESR2 and AHR/ARNT signalingpubmed:28263910
  • Antiestrogenic Activity of the Xi-Huang Formula for Breast Cancer by Targeting the Estrogen Receptor αPubmed:29975948
  • In Vitro Study of Mechanisms Underlying the Developmental Effects of Bisphenol A Using Human Fetal Lung Fibroblasts
  • Vitamin D receptor in breast cancer tissues and its relation to estrogen receptor alpha (ER-a) gene expression and serum 25-hydroxyvitamin D levels in Egyptian Doi: 10.1016/j.clbc.2018.12.019
  • ERα Gene Promoter Methylation in Cognitive Function and Quality of Life of Patients With Alzheimer DiseasePubmed: 30947592
  • The Effector Domain of the Influenza A Virus Nonstructural Protein NS1 Triggers Host Shutoff by Mediating Inhibition and Global Deregulation of Host Transcription …34488451
  • Modifying effect of obesity on the content of sex hormones and their receptors in endometrial adenocarcinoma and its surrounding tissue

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