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Mouse GHRL(Ghrelin) ELISA Kit

  • Cat.No.:E-EL-M0551

  • Reactivity: Mouse

To Purchase E-EL-M0551

Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $609
Qty:

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.
ItemSpecificationsStorage
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 1.20 3.80 0.50 1.30 3.50
Standard deviation 0.03 0.05 0.13 0.03 0.05 0.16
CV (%) 6.00 4.17 3.42 6.00 3.85 4.57

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) 90-105 97
EDTA plasma(n=8) 90-107 97
Cell culture media(n=8) 87-99 94

Linearity

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

Target Information

Database LinksSwissProt: Q9EQX0
SynonymsGHRL, MTLRP, Ghrelin
Research AreaCancer, Cardiovascular, Metabolism, Neuroscience, Signal transduction

Assay Procedures

elisa assay procedure 1

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

elisa assay procedure 2

2. Aspirate and wash the plate for 3 times

elisa assay procedure 3

3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

elisa assay procedure 4

4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

elisa assay procedure 5

5. Add 50μL Stop Solution

elisa assay procedure 6

6. Read the plate at 450nm immediately. Calculation of the results

Citations

  1. CELL (2023) IF: 66.849
    Light modulates glucose metabolism by a retina-hypothalamus-brown adipose tissue axis

    DOI: 10.1016/j.cell.2022.12.024

    PMID: 36669474

    Sample: Serum
  2. Gut Microbes (2021) IF: 10.245
    Activation of ectopic olfactory receptor 544 induces GLP-1 secretion and regulates gut inflammation

    DOI: 10.1080/19490976.2021.1987782

    PMID: 34674602

    Sample: plasma
  3. MOLECULAR NUTRITION & FOOD RESEARCH (2020) IF: 5.309
    Gymnaster Koraiensis Extract Alleviated Metabolic Syndrome Symptoms and Stimulated UCP1‐Independent Energy Consumption via AMPK Activation in White Adipose Tissue

    DOI: 10.1002/mnfr.202000490

    Sample: Serum
  4. ARCHIVES OF PHYSIOLOGY AND BIOCHEMISTRY (2021) IF: 4.076
    Asprosin, a novel glucogenic adipokine: a potential therapeutic implication in diabetes mellitus

    DOI: 10.1080/13813455.2021.1894178

    PMID: 33663304

    Sample: Serum
  5. Journal of Functional Foods (2021) IF: 4.451
    Heracleum dissectum Ledeb. ethanol extract attenuates metabolic syndrome symptoms in high-fat diet-induced obese mice by activating adiponectin/AMPK signaling

    DOI: 10.1016/j.jff.2021.104581

    Sample: blood
  6. TOXICOLOGY AND APPLIED PHARMACOLOGY (2020) IF: 3.347
    Role of TRPV1/TRPV3 channels in olanzapine-induced metabolic alteration: Possible involvement in hypothalamic energy-sensing, appetite regulation, inflammation and mesolimbic pathway

    DOI: 10.1016/j.taap.2020.115124

    PMID: 32652086

    Sample: Serum
  7. TOXICOLOGY AND APPLIED PHARMACOLOGY (2019) IF: 3.585
    Modeling of antipsychotic-induced metabolic alterations in mice: An experimental approach precluding psychosis as a predisposing factor

    DOI: 10.1016/j.taap.2019.114643

    PMID: 31254565

    Sample: Serum
  8. LIFE SCIENCES (2020) IF: 3.647
    Berberine attenuated olanzapine-induced metabolic alterations in mice: Targeting transient receptor potential vanilloid type 1 and 3 channels

    DOI: 10.1016/j.lfs.2020.117442

    Sample: serum
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