Multiplex Assay Kit for Ghrelin (GHRL) ,etc. by FLIA (Flow Luminescence Immunoassay)

MTLRP; Growth Hormone-Releasing Peptide

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Ghrelin (GHRL) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Ghrelin (GHRL) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 96-104 101
EDTA plasma(n=5) 91-105 95
heparin plasma(n=5) 97-104 101
sodium citrate plasma(n=5) 88-102 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Ghrelin (GHRL) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Ghrelin (GHRL) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 92-101% 89-96% 98-105% 95-103%
EDTA plasma(n=5) 84-92% 86-99% 79-103% 79-102%
heparin plasma(n=5) 83-102% 97-105% 91-98% 87-95%
sodium citrate plasma(n=5) 83-102% 81-96% 80-102% 88-105%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:GHRL) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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
Physiological and Histopathological Effects of Electroporation Pulse on Stomach of Rats
International Journal of Livestock Research Effect of Nutritional Supplementation on Treatment of Anoestrus Swamp Buffalo Cows and Heifers
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
npj Biofilms and Microbiomes The microbiota-gut-kidney axis mediates host osmoregulation in a small desert mammal Pubmed:35379849
Food Hydrocolloids Creating texturally tuneable, low calorie and palatable noodle-like food assemblies via microfluidics
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