ELISA Kit for Ghrelin (GHRL) Rattus norvegicus (Rat) Sandwich ELISA

MTLRP; Growth Hormone-Releasing Peptide

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  • ELISA Kit for Ghrelin (GHRL) Packages (Simulation)
  • ELISA Kit for Ghrelin (GHRL) Packages (Simulation)
  • ELISA Kit for Ghrelin (GHRL) Results demonstration
  • SEA991Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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) 92-101 96
EDTA plasma(n=5) 90-104 95
heparin plasma(n=5) 92-103 97

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) 98-105% 95-104% 83-96% 94-103%
EDTA plasma(n=5) 80-101% 93-102% 79-104% 89-96%
heparin plasma(n=5) 90-104% 97-105% 78-91% 86-98%

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

ELISA Kit for Ghrelin (GHRL)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Ghrelin (GHRL). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Ghrelin (GHRL). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Ghrelin (GHRL), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Ghrelin (GHRL) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Expression of ghrelin in human salivary glands and its levels in saliva and serum in Chinese obese children and adolescentsPubMed: 21075359
  • Presence of adropin, nesfatin-1, apelin-12, ghrelins and salusins peptides in the milk, cheese whey and plasma of dairy cowsScienceDirect: S0196978113000533
  • Postnatal administration of 2-oxoglutaric acid improves articular and growth plate cartilages and bone tissue morphology in pigs prenatally treated with dexamethasonePubMed: 23211309
  • The modifying effects of fish oil on fasting ghrelin mRNA expression in weaned ratsPubMed: 22842192
  • Changes in Ghrelin and Obestatin Levels before and after a Meal in Children with Simple Obesity and AnorexiaPubmed: 23774845
  • Gastric supply manipulation to modulate ghrelin production and enhance vascularization to the cardia: proof of the concept in a porcine modelPubmed:25294792
  • Embolization of Arterial Gastric Supply in Obesity (EMBARGO): an Endovascular Approach in the Management of Morbid Obesity. Proof of the Concept in the Porcine ModelPubmed:25511751
  • Effect of Sleeve Gastrectomy Plus Side-to-Side Jejunoileal Anastomosis for Type 2 Diabetes Control in an Obese Rat ModelPubMed: 26202420
  • Gastric Supply Manipulation to Modulate Ghrelin Production and Enhance Vascularization to the Cardia Proof of the Concept in a Porcine ModelPubMed: 25294792
  • Elevation of Fasting Ghrelin in Healthy Human Subjects Consuming a High-Salt Diet: A Novel Mechanism of Obesity?Pubmed:27240398
  • Effects of sleeve gastrectomy with jejuno-jejunal or jejuno-ileal loop onglycolipid metabolism in diabetic rats.pubmed:27621579
  • Increased serum ghrelin in preeclampsia: is ghrelin a friend or a foe?pubmed:27321099
  • The association between serum ghrelin levels and large-for-gestational-age fetuses in patients with gestational diabetes mellituspubmed:27791436
  • Effect of lifestyle improvement program on the biomarkersof adiposity, inflammation and gut hormones in overweight/obese Asian Indians with prediabetespubmed:28620678
  • Role of serotonin hormone in weight regain after sleeve gastrectomy pubmed:29228802
  • EFFECT OF Garcinia cambogia EXTRACT ON SERUM LEPTIN, GHRELIN, ADIPONECTIN AND INSULIN LEVELS AND BODY WEIGHTS IN RATS FED WITH HIGH …Slovetres:Source
  • The Relations Among Serum Ghrelin, Motilin and Gastric Emptying and Autonomic Function in Autoimmune GastritisPubmed:29753372
  • The Role of Umbilical Cord Blood Concentration of IGF-I, IGF-II, Leptin, Adiponectin, Ghrelin, Resistin, and Visfatin in Fetal GrowthPubmed: 30282106
  • The reparative effects of human adipose-derived mesenchymal stem cells in the chemotherapy damaged thymusPubmed: 30511904
  • Grape Seed Proanthocyanidin Affects Lipid Metabolism via Changing Gut Microflora and Enhancing Propionate Production in Weaned PigsPubmed: 31175811
  • Associations of apelin, leptin, irisin, ghrelin, insulin, glucose levels, and lipid parameters with physical activity during eight weeks of regular exercise trainingPubmed: 31290696
  • Ghrelin system is involved in improvements in glucose metabolism mediated by hyperbaric oxygen treatment in a streptozotocin‑induced type 1 diabetes mouse …Pubmed: 32901885
  • Omega-3 offers better hypothalamus protection by decreasing POMC expression and elevating ghrelin hormone: a prospective trial to overcome methotrexate …Pubmed: 32424681
  • Serum leptin level is associated with phase angle in CKD5 patients not undergoing dialysisPubmed: 30089154
  • Physiological and Histopathological Effects of Electroporation Pulse on Stomach of Rats
  • Effect of Nutritional Supplementation on Treatment of Anoestrus Swamp Buffalo Cows and Heifers
  • Relationship Between Orexin-A and Insulin Resistance in Patients with Type 2 Diabetes Mellitus
  • Altered Serum Marker of Adipokines Profile in Breast Cancer Women
  • The Association of Pre-Pregnancy BMI On Leptin, Ghrelin, Adiponectin, and IGF-1 In Breast Milk, A Case Control Study34348809
  • Nutritional Status, Selected Nutrients Intake and Their Relationship with the Concentration of Ghrelin and Adiponectin in Patients with Diabetic Nephropathy34959967
  • Effect of Sleeve Gastrectomy, Roux-En-Y Gastric Bypass, and Ileal Transposition on Myocardial Ischaemia–Reperfusion Injury in Non-Obese Non-Diabetic Rats34903800
  • The microbiota-gut-kidney axis mediates host osmoregulation in a small desert mammalPubmed:35379849
  • Creating texturally tuneable, low calorie and palatable noodle-like food assemblies via microfluidics

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