Multiplex Assay Kit for Glucagon Like Peptide 1 (GLP1) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Glucagon Like Peptide 1 (GLP1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Glucagon Like Peptide 1 (GLP1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Glucagon Like Peptide 1 (GLP1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Glucagon Like Peptide 1 (GLP1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 93-101 96
EDTA plasma(n=5) 89-102 93
heparin plasma(n=5) 87-96 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glucagon Like Peptide 1 (GLP1) ,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 Like Peptide 1 (GLP1) ,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 Like Peptide 1 (GLP1) ,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) 93-101% 92-101% 81-99% 80-103%
EDTA plasma(n=5) 81-101% 81-99% 82-101% 85-99%
heparin plasma(n=5) 80-95% 86-94% 83-102% 86-102%

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#:GLP1) 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
Diabetes, Obesity and Metabolism Specificity and sensitivity of commercially available assays for glucagon‐like peptide‐1 (GLP‐1): implications for GLP‐1 measurements in clinical studies Pubmed:25041349
Acta Histochemica Expression of cholecystokinin2-receptor in rat and human L cells and the stimulation of glucagon-like peptide-1 secretion by gastrin treatment Pubmed:25601282
Journal of Endocrinology Inhibitory effect of somatostatin on insulin secretion is not mediated via the CNS Pubmed:25630331
Obes Surg Effect of Sleeve Gastrectomy Plus Side-to-Side Jejunoileal Anastomosis for Type 2 Diabetes Control in an Obese Rat Model PubMed: 26202420
International Journal of Pharmacognosy and Phytochemical Research Anti-hiperglycemic Effect of Urena lobata Leaf Extract by Inhibition of Dipeptidyl Peptidase IV (DPP-IV) on Diabetic Rats Pdf: 7
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Int J Obes miR-200a reCavia (Guinea pig )lates Rheb-mediated amelioration of insulin resistance after duodenal-jejunal bypass Pubmed:27121251
Journal of Traditional and Complementary Medicine Incretin effect of Urena lobata leaves extract on structure and function of rats islet β-cells S2225411016302802
World Journal of Gastroenterology  Alterations in gut microbiota during remission and recurrence of diabetes after duodenal-jejunal bypass in rats pubmed:27547013
World Journal of Gastroenterology Effects of sleeve gastrectomy with jejuno-jejunal or jejuno-ileal loop onglycolipid metabolism in diabetic rats. pubmed:27621579
European Journal of Nutrition Improvement in glucose tolerance and insulin sensitivity by probiotic strains of Indian gut originin high-fat diet-fed C57BL/6J mice. pubmed:27757592
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Metabolism Deactivation of the NLRP3 inflammasome in infiltrating macrophages by duodenal-jejunal bypass surgery mediates improvement of beta cell function in type 2 diabetes 10.1016/j.metabol.2017.10.015
Obesity surgery The Effects of Duodenojejunal Omega Switch in Combination with High-Fat Diet and Control Diet on Incretins, Body Weight, and Glucose Tolerance in Sprague-Dawley Rats. pubmed:28840471
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Obesity Surgery  Preserving Duodenal-Jejunal (Foregut) Transit Does Not Impair Glucose Tolerance and Diabetes Remission Following Gastric Bypass in Type 2 Diabetes Sprague … Pubmed:29098544
Frontiers in Physiology Ileal Transposition Surgery Decreases Fat Mass and Improves Glucose Metabolism in Diabetic GK Rats: Possible Involvement of FGF21 Pubmed:29593555
Journal of Physiology and Biochemistry Downregulation of lncRNA MALAT1 contributes to renal functional improvement after duodenal-jejunal bypass in a diabetic rat model Pubmed:29781038
Metabolism Deactivation of the NLRP3 inflammasome in infiltrating macrophages by duodenal-jejunal bypass surgery mediates improvement of beta cell function in type 2 … Pubmed:29129820
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Peptides The influence of high fat diet on plasma incretins and insulin concentrations in Sprague-Dawley rats with diet-induced obesity and glucose intolerance undergoing … Doi: 10.1016/j.peptides.2019.04.001
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World J Diabetes Effect of oligofructose on resistance to postoperative high-fat diet-induced damage of metabolism in diabetic rats after sleeve gastrectomy 33889290
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