Multiplex Assay Kit for Fibroblast Growth Factor 15 (FGF15) ,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 Fibroblast Growth Factor 15 (FGF15) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Fibroblast Growth Factor 15 (FGF15) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Fibroblast Growth Factor 15 (FGF15) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Fibroblast Growth Factor 15 (FGF15) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 81-88 85
EDTA plasma(n=5) 83-101 94
heparin plasma(n=5) 95-102 99
sodium citrate plasma(n=5) 79-91 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Fibroblast Growth Factor 15 (FGF15) ,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 Fibroblast Growth Factor 15 (FGF15) ,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 Fibroblast Growth Factor 15 (FGF15) ,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) 94-104% 86-95% 83-104% 96-103%
EDTA plasma(n=5) 89-96% 85-102% 97-105% 99-105%
heparin plasma(n=5) 90-102% 79-93% 99-105% 80-96%
sodium citrate plasma(n=5) 89-96% 78-102% 90-103% 97-104%

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#:FGF15) 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 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Endocrinology. Naringenin Prevents Obesity, Hepatic Steatosis, and Glucose Intolerance in Male Mice Independent of Fibroblast Growth Factor 21 PubMed: 25774553
Inflammatory Bowel Diseases Alterations in Enterohepatic Fgf15 Signaling and Changes in Bile Acid Composition Depend on Localization of Murine Intestinal Inflammation pubmed:27580383
J Lipid Res. Commensal bacteria at the crossroad between cholesterol homeostasis and chronic inflammation in atherosclerosis. pubmed:28130274
BMC Endocrine Disorders The relationship between bile acid concentration, glucagon-like-peptide 1, fibroblast growth factor 15 and bile acid receptors in rats during progression of glucose intolerance 10.1186/s12902-017-0211-5
Journal of Physiology and Biochemistry The ileal FGF15/19 to hepatic FGFR4 axis regulates liver regeneration after partial hepatectomy in mice Pubmed:29468415
Developmental Cell FGF15 Activates Hippo Signaling to Suppress Bile Acid Metabolism and Liver Tumorigenesis Pubmed: 30745141
Endocrinology Interactions between the gravitostat and the fibroblast growth factor system for the regulation of body weight
Food & Function Lactoferrin promotes bile acid metabolism and reduces hepatic cholesterol deposition by inhibiting the farnesoid X receptor (FXR)-mediated enterohepatic axis Pubmed: 31626262
Journal of Translational Medicine Gut microbiota from coronary artery disease patients contributes to vascular dysfunction in mice by regulating bile acid metabolism and immune activation Pubmed: 33036625
Chin Med Si-Wu-Tang ameliorates fibrotic liver injury via modulating intestinal microbiota and bile acid homeostasis 34736501
Int J Biol Sci Dynamics of the gut-liver axis in rats with varying fibrosis severity Pubmed:35637968
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