RayBio® Human/Mouse/Rat Ghrelin EIA Kit optimized for serum and cell culture medium. Competition-based ELISA on a 96-well strip plate.

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Catalog #:
EIA-GHR
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Product Description

Specifications

Size 1 Plate Kit, 2 Plate Kit, 5 Plate Kit
Species Human, Mouse, Rat
Accession Number Q9UBU3
Gene Id 51738
Gene Symbols GHRL|MTLRP|UNQ524/PRO1066
Protein Name / Synonyms Appetite-regulating hormone (Growth hormone secretagogue) (Growth hormone-releasing peptide) (Motilin-related peptide) (Protein M46) [Cleaved into: Ghrelin-27 Ghrelin-28 (Ghrelin) Obestatin]
Quantitative/Semi-Quantitative Quantitative
Specificity Cross Reactivity: This EIA kit shows no cross-reactivity with any of the cytokines tested: Nesfatin, Angiotensin II, NPY and APC.
Compatible Sample Types Cell Culture Supernatants, Serum
Solid Support 96-well Microplate
Method Of Detection Colorimetric
Design Principle Competition-based
Sensitivity 161 pg/ml or 12.46 pM
Detection Range 0.1-1,000 ng/ml
Estimated Lead Time 1-2 business days
Shipping Type Blue ice
Storage -20°C

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Introduction

Obesity, which is characterized by excessive accumulation of adipose tissue in the body, has become one of the greatest public health challenges. Obesity is not only associated with health problems linked to increased weight-dependent pressure overload on lung, joints and bones, but also a important risk factor for life-threatening diseases such as cardiovascular diseases, type 2 diabetes and certain cancers.

Ghrelin is synthesized as a preprohormone, and then proteolytically processed to yield a 28-amino acid peptide. Synthesis of ghrelin occurs predominantly in epithelial cells lining the fundus of the stomach, with smaller amounts produced in the placenta, kidney, pituitary and hypothalamus.

Ghrelin has emerged as the first circulating hunger hormone. Ghrelin increases food intake and thus fat mass by an action exerted at the level of the hypothalamus. They activate cells in the arcuate nucleus that include the orexigenic neuropeptide Y (NPY) neurons. Ghrelin-responsiveness of these neurons is both leptin and insulin sensitive. Ghrelin also activates the mesolimbic cholinergic-dopaminergic reward link, a circuit that communicates the hedonic and reinforcing aspects of natural rewards, such as food.

Ghrelin levels in the plasma of obese individuals are lower than those in leaner individuals except in the case of Prader-Willi syndrome-induced obesity. Those suffering from the eating disorder anorexia nervosa have high plasma levels of ghrelin compared to both the constitutionally thin and normal-weight controls. These findings suggest that ghrelin plays a role in both anorexia and obesity. Ghrelin levels are also high in patients who have cancer-induced cachexia.

Product Features

  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Standard Peptide
  • Assay Diluent(s)
  • Biotinylated Peptide
  • HRP-Streptavidin
  • TMB One-Step Substrate
  • Stop Solution
  • Assay Diagram
  • Positive Control Sample
  • Capture Antibody
  • User Manual
Other Materials Required
  • Distilled or deionized water
  • Precision pipettes to deliver 2 µl to 1 ml volumes
  • Adjustable 1-25 ml pipettes for reagent preparation
  • 100 ml and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Orbital shaker
  • Aluminum foil
  • Saran Wrap
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • SigmaPlot software (or other software that can perform four-parameter logistic regression models)
Protocol Outline
  • Prepare all reagents, samples and standards as instructed.
  • Add 100 µl detection antibody to each well.
  • Incubate 1.5 h at RT or O/N at 4°C.
  • Add 100 µl standard or sample to each well.
  • Incubate 2.5 h at RT.
  • Add 100 µl prepared streptavidin solution.
  • Incubate 45 min at RT.
  • Add 100 µl TMB One-Step Substrate Reagent to each well.
  • Incubate 30 min at RT.
  • Add 50 µl Stop Solution to each well.
  • Read plate at 450 nm immediately.
  • Storage/Stability

    Standard, Biotinylated Ghrelin (GHR) peptide, and Positive Control should be stored at -20°C after arrival. Avoid multiple freeze-thaws. The remaining kit components may be stored at 4°C. Opened Microplate Wells and antibody (Item N) may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack and reseal along entire edge.
    Dang R., Jiang P., Cai H., et al. Vitamin D deficiency exacerbates atypical antipsychotic-induced metabolic side effects in rats: involvement of the INSIG/SREBP pathway. Eur Neuropsychopharmacol. 2015 Aug;25(8):1239-47. doi: 10.1016/j.euroneuro.2015.04.028.
    Species:
    Rat
    Sample Type:
    Serum (Vitamin D deficient Rat model on pyschotic states)
    Ricken, Roland, et al. "Ghrelin serum concentrations are associated with treatment response during lithium augmentation of antidepressants." International Journal of Neuropsychopharmacology 20.9 (2017): 692-697.
    Species:
    Human
    Sample Type:
    Serum
    Hehir, Mark P., et al. "Ghrelin concentrations in maternal and cord blood of Type 1 diabetic and non-diabetic pregnancies at term." Endocrine (2013): 1-3.
    Species:
    Human
    Sample Type:
    Serum (maternal and fetal ghrelin in GD vs control pregnancy)
    Wysokinski A., Kowalski M., Kloszewska I. Serum levels of desacyl ghrelin in patients with schizophrenia on clozapine monotherapy Accepted manuscript online: 28 APR 2014 09:37PM EST | DOI: 10.1111/pcn.12199 [Epub Ahead of Print]
    Species:
    Human
    Sample Type:
    Serum (Human Serum)
    Russo, Francesco, et al. "The effects of fluorouracil, epirubicin, and cyclophosphamide (FEC60) on the intestinal barrier function and gut peptides in breast cancer patients: an observational study." BMC cancer 13.1 (2013): 56.
    Species:
    Human
    Sample Type:
    Serum (intestinal barrier function assessment during chemotherapy in breast cancer patients)
    Han J., Kim H., Lee J., Choi M., Kim Y., Son C. Repeated Sense of Hunger Leads to the Development of Visceral Obesity and Metabolic Syndrome in a Mouse Model . PLOS One Published: May 30, 2014. DOI: 10.1371/journal.pone.0098276
    Species:
    Rat
    Sample Type:
    Serum
    Plum L., et al. The Obesity Susceptibility Gene Carboxypeptidase E Links FoxO1 Signaling in Hypothalamic Pro-opiomelanocortin Neurons with Regulation of Food Intake. Nat Med. 2009 Oct;15(10):1195-201. doi: 10.1038/nm.2026
    Species:
    Mouse
    Sample Type:
    Serum
    Polat Z., et al. Plasma Ghrelin Levels in Patients with Familial Mediterranean Fever. Digestive Diseases and Sciences. June 2012, Volume 57, Issue 6, pp 1660-1663.
    Species:
    Human
    Sample Type:
    Serum
    Cobanoglu N., Dalkan C., Galip N., et al. Is calprotectin a marker of tobacco smoke related inflammation?: a pilot study in children. Inhal Toxicol. 2012 Jul;24(8):486-91. doi: 10.3109/08958378.2012.693137.
    Species:
    Human
    Sample Type:
    Serum (Tobacco smoke related)
    Polat Z., Kilciler G., Ozel A., et al. Plasma ghrelin levels in patients with familial Mediterranean fever. Dig Dis Sci. 2012 Jun;57(6):1660-3. doi: 10.1007/s10620-012-2049-z.
    Species:
    Human
    Sample Type:
    Serum (Familial Mediterranean Fever)
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