Multiplex Assay Kit for Estrogen Receptor Alpha (ERa) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Estrogen Receptor Alpha (ERa) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Estrogen Receptor Alpha (ERa) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 96-105 101
EDTA plasma(n=5) 97-104 101
heparin plasma(n=5) 92-101 96
sodium citrate plasma(n=5) 88-98 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Estrogen Receptor Alpha (ERa) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 83-93% 80-98% 82-104% 90-98%
EDTA plasma(n=5) 78-105% 95-102% 78-88% 82-91%
heparin plasma(n=5) 98-105% 89-98% 93-101% 86-95%
sodium citrate plasma(n=5) 87-95% 88-95% 85-102% 82-98%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:ERa) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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
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
cellular microbiology Influenza A virus NS1 protein-induced JNK activation and apoptosis are not functionally linked. pubmed:28076660
Journal of Steroid Biochemistry and Molecular Biology 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 signaling pubmed:28263910
Cellular Physiology and Biochemistry Antiestrogenic Activity of the Xi-Huang Formula for Breast Cancer by Targeting the Estrogen Receptor α Pubmed:29975948
Carleton University Research Virtual Environment In Vitro Study of Mechanisms Underlying the Developmental Effects of Bisphenol A Using Human Fetal Lung Fibroblasts
Clinical Breast Cancer 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
Journal of geriatric psychiatry and neurology ERα Gene Promoter Methylation in Cognitive Function and Quality of Life of Patients With Alzheimer Disease Pubmed: 30947592
mBio The Effector Domain of the Influenza A Virus Nonstructural Protein NS1 Triggers Host Shutoff by Mediating Inhibition and Global Deregulation of Host Transcription … 34488451
Cardiometry Modifying effect of obesity on the content of sex hormones and their receptors in endometrial adenocarcinoma and its surrounding tissue
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