ELISA Kit for G Protein Coupled Estrogen Receptor 1 (GPER)

GPR30; CMKRL2; DRY12; FEG-1; GPCR-Br; LERGU; LyGPR; Membrane estrogen receptor; Chemoattractant receptor-like 2; Flow-induced endothelial G-protein coupled receptor 1

Specificity

This assay has high sensitivity and excellent specificity for detection of G Protein Coupled Estrogen Receptor 1 (GPER).
No significant cross-reactivity or interference between G Protein Coupled Estrogen Receptor 1 (GPER) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant G Protein Coupled Estrogen Receptor 1 (GPER) and the recovery rates were calculated by comparing the measured value to the expected amount of G Protein Coupled Estrogen Receptor 1 (GPER) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-99 88
EDTA plasma(n=5) 95-102 98
heparin plasma(n=5) 88-96 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level G Protein Coupled Estrogen Receptor 1 (GPER) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level G Protein Coupled Estrogen Receptor 1 (GPER) 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 G Protein Coupled Estrogen Receptor 1 (GPER) 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) 93-101% 95-103% 92-99% 80-102%
EDTA plasma(n=5) 84-103% 96-103% 93-101% 90-98%
heparin plasma(n=5) 93-101% 86-97% 92-99% 82-99%

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
Molecular and Cellular Endocrinology Isomer-nonspecific action of dichlorodiphenyltrichloroethane on aryl hydrocarbon receptor and G-protein-coupled receptor 30 intracellular signaling in apoptotic neuronal cells Pubmed:24859647
J Steroid Biochem Mol Biol Neuroprotective action of raloxifene against hypoxia-induced damage in mouse hippocampal cells depends on ERα but not ERβ or GPR32 signalling PubMed: 24846831
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
Psychiatry research Serum levels of G protein-coupled estrogen receptor 1 (GPER1) in drug-naive patients with generalized anxiety disorder. pubmed:27512921
Clinical Psychopharmacology Neuroscience. Increased Serum G Protein-coupled Estrogen Receptor 1 Levels and Its Diagnostic Value in Drug Naïve Patients with Major Depressive Disorder pubmed:29073745
Psychiatry Research Decreased levels of G protein-coupled estrogen receptor in children with autism spectrum disorders. pubmed:28734238
Neuropsychiatric disease and treatment serum levels of gPer-1 in euthymic bipolar patients Pubmed:29618927
European archives of oto-rhino-laryngology An evaluation of G-protein coupled membrane estrogen receptor-1 level in stuttering Pubmed:29299745
Bosnian Journal of Basic Medical Sciences Evaluation of estrogen and G protein-coupled estrogen receptor 1 (GPER) levels in drug-naïve patients with attention deficit hyperactivity disorder (ADHD) Pubmed:29659348
Carleton University Research Virtual Environment In Vitro Study of Mechanisms Underlying the Developmental Effects of Bisphenol A Using Human Fetal Lung Fibroblasts
Am J Otolaryngol GPER-1 and sex-hormone levels in patients with otosclerosis Pubmed: 32144019
Andrologia. Relationship between the G protein–coupled oestrogen receptor and spermatogenesis, and its correlation with male infertility Pubmed: 32776559
Journal of Radiation Research and Applied Sciences GPER-1 in chronic hepatitis B
FASEB J Endocrine disruption in Crohn's disease: Bisphenol A enhances systemic inflammatory response in patients with gut barrier translocation of dysbiotic microbiota?¡­ 34085740
Comparing the Impact of Serum GPER-1 and Oxidant/Antioxidant Levels on Retinopathy in Diabetic Patients and Healthy Individuals: A Pilot Study
Horm Mol Biol Clin Investig The change pattern in serum G protein-coupled estrogen receptor-1 (GPER1) levels during pregnancy with and without gestational diabetes mellitus 34787384
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