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Rat GHRL(Ghrelin) ELISA Kit

Cat.No.:E-EL-R0842
Reactivity: Rat

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96T
48T
24T
96T*5
96T*10

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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
Assay time	2.0h
Detection range	0.16-10 ng/mL
Sensitivity	0.10 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes GHRL in samples. No significant cross-reactivity or interference between GHRL 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 GHRL concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with GHRL. During the reaction, GHRL in the sample or standard competes with a fixed amount of GHRL on the solid phase supporter for sites on the Biotinylated Detection Ab specific to GHRL. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of GHRL in tested samples can be calculated 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℃
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 GHRL were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level GHRL 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)	0.50	0.90	3.80	0.50	0.90	4.00
Standard deviation	0.03	0.05	0.13	0.03	0.04	0.20
CV (%)	6.00	5.56	3.42	6.00	4.44	5.00

Recovery

The recovery of GHRL 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)	92-103	97
EDTA plasma(n=8)	85-101	92
Cell culture media(n=8)	93-106	98

Linearity

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

Target Information

Synonyms	Obestatin Preprohormone, MTLRP, Ghrelin
Research Area	Cancer, Cardiovascular, Metabolism, Neuroscience, Signal transduction

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. 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
Food & Function (2022) IF: 5.396
Lactiplantibacillus plantarum CCFM1019 attenuate polycystic ovary syndrome through butyrate dependent gut–brain mechanism

DOI: 10.1039/D1FO01744F

PMID: 35044398
Sample: serum
Scientific Reports (2021) IF: 4.38
Acetate rescues defective brain-adipose metabolic network in obese Wistar rats by modulation of peroxisome proliferator-activated receptor-γ

DOI: 10.1038/s41598-021-98605-5

PMID: 34556775
Sample: plasma
PSYCHONEUROENDOCRINOLOGY (2022) IF: 4.693
Increased growth hormone secretagogue receptor-1a (GHSR-1a) in hypothalamus during olanzapine treatment in rats

DOI: 10.1016/j.psyneuen.2022.105862

PMID: 35835020
Sample: Serum
MOLECULES (2018) IF: 3.098
The Effects of Plant-Derived Oleanolic Acid on Selected Parameters of Glucose Homeostasis in a Diet-Induced Pre-Diabetic Rat Model

DOI: 10.3390/molecules23040794

Sample: plasma
Scientific Reports (2020) IF: 3.998
Effect of different levels of hydrolysable tannin intake on the reproductive hormones and serum biochemical indices in healthy female rats

DOI: 10.1038/s41598-020-77672-0

PMID: 33244120
Sample: serum
ARCHIVES OF PHYSIOLOGY AND BIOCHEMISTRY (2019) IF: 2.11
Evaluation of the effects of bredemolic acid on selected markers of glucose homeostasis in diet-induced prediabetic rats

DOI: 10.1080/13813455.2019.1680697

Sample: plasma
NEUROSCIENCE LETTERS (2017) IF: 2.18
The effects of intracerebroventricular infusion of irisin on feeding behaviour in rats

DOI: 10.1016/j.neulet.2017.02.066

Sample: Serum
Integrative Medicine Research (2019) IF: 2.379
Orthosiphon stamineus as a potential antidiabetic drug in maternal hyperglycemia in streptozotocin-induced diabetic rats

DOI: 10.1016/j.imr.2019.05.006

Sample: plasm
Open Life Sciences (2022) IF: 1.311
Physical activity enhances fecal lactobacilli in rats chronically drinking sweetened cola beverage

DOI: 10.1515/biol-2022-0070

Sample: plasma
BIOLOGIA (2022) IF: 1.653
The effects of β-escin on inflammation, oxidative stress and Langerhans islet cells in high-fat diet and streptozotocin injection induced experimental type-2 diabetes model

DOI: 10.1007/s11756-022-01266-6

Sample: serum
BIOTECHNIC & HISTOCHEMISTRY (2021) IF: 1.718
Short term effects of experimental gastric outlet obstruction and truncal vagotomy on gut hormones

DOI: 10.1080/10520295.2021.1896780

PMID: 33722110
Sample: serum
Vascular Health and Risk Management (2023)
The Effectiveness of Nigella sativa and Ginger as Appetite Suppressants: An Experimental Study on Healthy Wistar Rats

DOI: 10.2147/VHRM.S396295

Sample: serum
Scientific African (2021)
Effect of Allium sativum extract in combination -with orlistat on insulin resistance and disrupted metabolic hormones in high fat diet induced obese rats

DOI: 10.1016/j.sciaf.2021.e00994

Sample: serum

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