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Rat GnRHR(Gonadotropin-Releasing Hormone Receptor) ELISA Kit

Cat.No.:E-EL-R0450
Reactivity: Rat

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96T
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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.31-20 ng/mL
Sensitivity	0.19 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Rat GnRHR in samples. No significant cross-reactivity or interference between Rat GnRHR and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Rat GnRHR concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Rat GnRHR. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat GnRHR and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat GnRHR, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Rat GnRHR. You can calculate the concentration of Rat GnRHR in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat GnRHR were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat GnRHR were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	1.03	2.54	8.80	1.08	2.42	9.10
Standard deviation	0.06	0.12	0.28	0.07	0.10	0.48
CV (%)	5.83	4.72	3.18	6.48	4.13	5.27

Recovery

The recovery of Rat GnRHR spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	84-98	91
EDTA plasma (n=8)	90-107	98
Cell culture media (n=8)	87-101	92

Linearity

Samples were spiked with high concentrations of Rat GnRHR and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Research Area

Cancer, Neuroscience

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

CARBOHYDRATE POLYMERS (2022) IF: 9.381
Butylated starch alleviates polycystic ovary syndrome by stimulating the secretion of peptide tyrosine-tyrosine and regulating faecal microbiota

DOI: 10.1016/j.carbpol.2022.119304

PMID: 35422283
Sample: serum
ENVIRONMENTAL POLLUTION (2022) IF: 9.988
Acute exposure to microcystins affects hypothalamic-pituitary axes of male rats

DOI: 10.1016/j.envpol.2022.120843

Sample: serum
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY (2022) IF: 7.129
Arsenic exposure diminishes ovarian follicular reserve and induces abnormal steroidogenesis by DNA methylation

DOI: 10.1016/j.ecoenv.2022.113816

PMID: 36068745
Sample: Serum
Scientific Reports (2022) IF: 4.38
Testicular ultrastructure and hormonal changes following administration of tenofovir disoproxil fumarate-loaded silver nanoparticle in type-2 diabetic rats

DOI: 10.1038/s41598-022-13321-y

PMID: 35688844
Sample: serum
Scientific Reports (2020) IF: 3.998
Reproductive effects of subchronic exposure to acetamiprid in male rats

DOI: 10.1038/s41598-020-65887-0

PMID: 32488017
Sample: Plasma,Tissue homogenate

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