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) 82-102 91
EDTA plasma(n=5) 85-99 92
heparin plasma(n=5) 95-103 99
sodium citrate plasma(n=5) 81-101 90

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) 80-93% 89-98% 98-105% 89-97%
EDTA plasma(n=5) 86-97% 90-104% 84-95% 91-98%
heparin plasma(n=5) 78-104% 79-89% 95-105% 80-99%
sodium citrate plasma(n=5) 94-101% 79-97% 80-102% 93-104%

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
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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
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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
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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
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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
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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
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