ELISA Kit for Ghrelin (GHRL)

MTLRP; Growth Hormone-Releasing Peptide

Specificity

This assay has high sensitivity and excellent specificity for detection of Ghrelin (GHRL).
No significant cross-reactivity or interference between Ghrelin (GHRL) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Ghrelin (GHRL) and the recovery rates were calculated by comparing the measured value to the expected amount of Ghrelin (GHRL) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-104 98
EDTA plasma(n=5) 96-103 101
heparin plasma(n=5) 80-97 84

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Ghrelin (GHRL) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Ghrelin (GHRL) 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 Ghrelin (GHRL) 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) 82-102% 88-102% 82-101% 80-90%
EDTA plasma(n=5) 88-97% 97-105% 86-103% 79-91%
heparin plasma(n=5) 78-91% 80-88% 79-96% 78-101%

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 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Archives of Oral Biology Expression of ghrelin in human salivary glands and its levels in saliva and serum in Chinese obese children and adolescents PubMed: 21075359
Department of Medical Biochemistry and Clinical Biochemistry Presence of adropin, nesfatin-1, apelin-12, ghrelins and salusins peptides in the milk, cheese whey and plasma of dairy cows ScienceDirect: S0196978113000533
Journal of Physiology and Pharmacology Postnatal administration of 2-oxoglutaric acid improves articular and growth plate cartilages and bone tissue morphology in pigs prenatally treated with dexamethasone PubMed: 23211309
Gene The modifying effects of fish oil on fasting ghrelin mRNA expression in weaned rats PubMed: 22842192
Hormone Research in Paediatrics Changes in Ghrelin and Obestatin Levels before and after a Meal in Children with Simple Obesity and Anorexia Pubmed: 23774845
Surgical Innovation Gastric supply manipulation to modulate ghrelin production and enhance vascularization to the cardia: proof of the concept in a porcine model Pubmed:25294792
Obesity Surgery Embolization of Arterial Gastric Supply in Obesity (EMBARGO): an Endovascular Approach in the Management of Morbid Obesity. Proof of the Concept in the Porcine Model Pubmed:25511751
Obes Surg Effect of Sleeve Gastrectomy Plus Side-to-Side Jejunoileal Anastomosis for Type 2 Diabetes Control in an Obese Rat Model PubMed: 26202420
Surg Innov Gastric Supply Manipulation to Modulate Ghrelin Production and Enhance Vascularization to the Cardia Proof of the Concept in a Porcine Model PubMed: 25294792
Nutrients.  Elevation of Fasting Ghrelin in Healthy Human Subjects Consuming a High-Salt Diet: A Novel Mechanism of Obesity? Pubmed:27240398
World Journal of Gastroenterology Effects of sleeve gastrectomy with jejuno-jejunal or jejuno-ileal loop onglycolipid metabolism in diabetic rats. pubmed:27621579
Ginekologia Polska Increased serum ghrelin in preeclampsia: is ghrelin a friend or a foe? pubmed:27321099
Gynecological Endocrinology The association between serum ghrelin levels and large-for-gestational-age fetuses in patients with gestational diabetes mellitus pubmed:27791436
Acta Diabetologica Effect of lifestyle improvement program on the biomarkersof adiposity, inflammation and gut hormones in overweight/obese Asian Indians with prediabetes pubmed:28620678
Scandinavian Journal of Clinical and Laboratory Investigation Role of serotonin hormone in weight regain after sleeve gastrectomy pubmed:29228802
Slovenian Veterinary Research EFFECT OF Garcinia cambogia EXTRACT ON SERUM LEPTIN, GHRELIN, ADIPONECTIN AND INSULIN LEVELS AND BODY WEIGHTS IN RATS FED WITH HIGH … Slovetres:Source
The American Journal of the Medical Sciences The Relations Among Serum Ghrelin, Motilin and Gastric Emptying and Autonomic Function in Autoimmune Gastritis Pubmed:29753372
American Journal of Perinatology The Role of Umbilical Cord Blood Concentration of IGF-I, IGF-II, Leptin, Adiponectin, Ghrelin, Resistin, and Visfatin in Fetal Growth Pubmed: 30282106
Stem Cells and Development The reparative effects of human adipose-derived mesenchymal stem cells in the chemotherapy damaged thymus Pubmed: 30511904
The Journal of Nutrition Grape Seed Proanthocyanidin Affects Lipid Metabolism via Changing Gut Microflora and Enhancing Propionate Production in Weaned Pigs Pubmed: 31175811
Arch Physiol Biochem Associations of apelin, leptin, irisin, ghrelin, insulin, glucose levels, and lipid parameters with physical activity during eight weeks of regular exercise training Pubmed: 31290696
Mol Med Rep Ghrelin system is involved in improvements in glucose metabolism mediated by hyperbaric oxygen treatment in a streptozotocin‑induced type 1 diabetes mouse … Pubmed: 32901885
endocrine Omega-3 offers better hypothalamus protection by decreasing POMC expression and elevating ghrelin hormone: a prospective trial to overcome methotrexate … Pubmed: 32424681
plos one Serum leptin level is associated with phase angle in CKD5 patients not undergoing dialysis Pubmed: 30089154
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iraqi journal of science Relationship Between Orexin-A and Insulin Resistance in Patients with Type 2 Diabetes Mellitus
Indian Journal of Forensic Medicine & Toxicology Altered Serum Marker of Adipokines Profile in Breast Cancer Women
Br J Nutr The Association of Pre-Pregnancy BMI On Leptin, Ghrelin, Adiponectin, and IGF-1 In Breast Milk, A Case Control Study 34348809
Nutrients Nutritional Status, Selected Nutrients Intake and Their Relationship with the Concentration of Ghrelin and Adiponectin in Patients with Diabetic Nephropathy 34959967
scientific reports Effect of Sleeve Gastrectomy, Roux-En-Y Gastric Bypass, and Ileal Transposition on Myocardial Ischaemia–Reperfusion Injury in Non-Obese Non-Diabetic Rats 34903800
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