ELISA Kit for Estrogen Receptor Beta (ERb)

ER-B; ESRB; ESTRB; NR3A2; NR3-A2; ESR2; Estrogen Receptor 2; Nuclear Receptor Subfamily 3,Group A,Member 2

Specificity

This assay has high sensitivity and excellent specificity for detection of Estrogen Receptor Beta (ERb).
No significant cross-reactivity or interference between Estrogen Receptor Beta (ERb) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 96-105 102
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 86-95 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Estrogen Receptor Beta (ERb) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Estrogen Receptor Beta (ERb) 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 Beta (ERb) 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) 88-98% 91-105% 80-101% 87-104%
EDTA plasma(n=5) 81-103% 96-105% 79-102% 87-94%
heparin plasma(n=5) 85-99% 95-103% 78-102% 79-96%

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.

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Magazine Citations
The Journal of Steroid Biochemistry and Molecular Biology Neuroprotective action of raloxifene against hypoxia-induced damage in mouse hippocampal cells depends on ERα but not ERβ or GPR30 signalling Pubmed:24846829
Biochem Biophys Res Commun Differential expression of estrogen receptor α and β isoforms in multiple and solitary leiomyomas PubMed: 26529545
Mol Neurobiol Selective Aryl Hydrocarbon Receptor Modulator 3,3′-Diindolylmethane Impairs AhR and ARNT Signaling and Protects Mouse Neuronal Cells Against Hypoxia PubMed: 26476840
J Steroid Biochem Mol Biol Neuroprotective action of raloxifene against hypoxia-induced damage in mouse hippocampal cells depends on ERα but not ERβ or GPR31 signalling PubMed: 24846830
Journal of Environmental Sciences Bisphenol A exposure alters release of immune and developmental modulators and expression of estrogen receptors in human fetal lung fibroblasts science:S1001074216300924
Biol Open. Involvement of BDNF/TrkB and ERK/CREB axes in nitroglycerin-induced rat migraine and effects of estrogen on these signals in the migraine. pubmed:27875242
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