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Multiplex Assay Kit for Glucagon (GCG) ,etc. by FLIA (Flow Luminescence Immunoassay)

GC; GLP1; GLP2; GRPP; Glicentin-Related Polypeptide; Glucagen; Oxyntomodulin; Incretin hormone

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 78-96 80
EDTA plasma(n=5) 98-105 101
heparin plasma(n=5) 79-97 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glucagon (GCG) ,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 Glucagon (GCG) ,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 Glucagon (GCG) ,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) 81-101% 88-102% 97-105% 78-98%
EDTA plasma(n=5) 78-94% 85-97% 80-105% 79-98%
heparin plasma(n=5) 95-103% 79-88% 96-104% 89-97%

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#:GCG) 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
Molecular Metabolism PER2 promotes glucose storage to liver glycogen during feeding and acute fasting by inducing Gys2 PTG and GL expression PubMed: PMC3773838
Exp Biol Med (Maywood). Increasing glucagon secretion could antagonize the action of exogenous insulin for glycemic control in streptozocin-induced diabetic rhesus monkeys. Pubmed: 23760004
European Journal of Endocrinology Specificity and sensitivity of commercially available assays for glucagon and oxyntomodulin measurement in human Eje-online: Source
American Diabetes Association IL-21 is a major negative regulator of IRF4-dependent lipolysis affecting Tregs in adipose tissue and systemic insulin sensitivity Pubmed:24430438
Journal of Pharmacy and Pharmacology Enzogenol improves diabetes-related metabolic change in C57BL/KsJ-db/db?mice, a model of type 2 diabetes mellitus Pubmed:24611864
European Journal of Pharmacology Subcutaneous administration of liraglutide ameliorates learning and memory impairment by modulating tau hyperphosphorylation via the glycogen synthase kinase-3尾 pathway in an amyloid 尾 protein induced alzheimer disease mouse model Pubmed:27131827
Appl Physiol Nutr Metab The glucoreCavia (Guinea pig )latory response to high-intensity aerobic exercise following training in rats with insulin-treated type 1 diabetes mellitus Pubmed:27175938
American Journal of Translational Research Subcutaneous liraglutide ameliorates methylglyoxal-induced Alzheimer-like tau pathology and cognitive impairment by modulating tau hyperphosphorylation and glycogen synthase kinase-3β. pubmed:28337257
Biomedicine & Pharmacotherapy Vitamin D3 intake as regulator of insulin degrading enzyme and insulin receptor phosphorylationin diabetic rats. pubmed:27930980
Diabetologia Ginsenoside Rg5 attenuates hepatic glucagon response via suppression of succinate-associated HIF-1α induction in HFD-fed mice pubmed:28280902
International Journal of Molecular Sciences Effects of Acute Cold Stress on Liver O-GlcNAcylation and Glycometabolism in Mice Pubmed: 30231545
European Journal of Pharmacology Exogenous obestatin decreases beta-cell apoptosis and alfa-cell proliferation in high fat diet and streptozotocin induced type 2 diabetic rats Pubmed: 30776368
Stress Regulating glycolysis, the TLR4 signal pathway and expression of RBM3 in mouse liver in response to acute cold exposure Pubmed: 30821572
international journal of molecular sciences Effects of Cold-inducible RNA-binding Protein (CIRP) on Liver Glycolysis during Acute Cold Exposure in C57BL/6 Mice Pubmed: 30909542
Diabetes-Metabolism Research and Reviews Liraglutide Combined with Human Umbilical Cord Mesenchymal Stem Cell Transplantation inhibits beta‐cells apoptosis via mediating the ASK1/JNK/BAX pathway in Pubmed: 31411368
CELL DISCOVERY In vitro expansion of pancreatic islet clusters facilitated by hormones and chemicals Pubmed: 32284878
endocrine journal Mice with nucleus tractus solitarius injury induced by chronic restraint stress present impaired ability to raise blood glucose and glucagon levels when blood glucose … Pubmed: 32249244
Frontiers in Endocrinology RFRP-3, the Mammalian Ortholog of GnIH, Is a Novel Modulator Involved in Food Intake and Glucose Homeostasis Pubmed: 32328034
Molecular Medicine Reports Acarbose ameliorates spontaneous type‑2 diabetes in db/db mice by inhibiting PDX‑1 methylation Pubmed: 33236139
CHEMICO-BIOLOGICAL INTERACTIONS Modulatory role of imatinib mesylate on pancreatic β-cells' secretory functions in an STZ rat model of diabetes mellitus Pubmed: 32710900
Mol Cell Endocrinol Mof acetyltransferase inhibition ameliorates glucose intolerance and islet dysfunction of type 2 diabetes via targeting pancreatic α-cells 34391847
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