Multiplex Assay Kit for Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) ,etc. by FLIA (Flow Luminescence Immunoassay)

NTN; ABCD3; C3Xkine; CXC3; CXC3C; NTT; SCYD1; ABCD3; FKN; Neurotactin; Fractalkine; Small Inducible Cytokine Subfamily D(Cys-X3-Cys)Member 1

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 91-105 98
EDTA plasma(n=5) 81-88 84
heparin plasma(n=5) 78-98 81

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) ,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 Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) ,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 Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) ,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) 96-103% 91-104% 78-99% 79-98%
EDTA plasma(n=5) 80-101% 89-97% 91-99% 80-92%
heparin plasma(n=5) 96-104% 83-94% 95-103% 97-105%

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#:CX3CL1) 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
World Journal of Gastroenterology Preliminary study correlating CX3CL1/CX3CR1 expression with gastric carcinoma and gastric carcinoma perineural invasion NCBI: PMC3989981
International Journal of Colorectal Disease Downregulation of CX3CR1 ameliorates experimental colitis: evidence for CX3CL1-CX3CR1-mediated immune cell recruitment. pubmed:27942903
Frontiers in Cellular and Infection Microbiology Reduced CX3CL1 Secretion Contributes to the Susceptibility of Oral Leukoplakia-Associated Fibroblasts to Candida albicans. pubmed:27891323
Blood Coagul Fibrinolysis Low shear stress upregulates the expression of fractalkine through the activation of mitogen-activated protein kinases in endothelial cells Pubmed:29406386
Journal of Neuroinflammation The prenatal challenge with lipopolysaccharide and polyinosinic: polycytidylic acid disrupts CX3CL1-CX3CR1 and CD200-CD200R signalling in the brains of … Pubmed: 32829711
Cells Maternal Immune Activation Sensitizes Male Offspring Rats to Lipopolysaccharide-Induced Microglial Deficits Involving the Dysfunction of CD200–CD200R and … Pubmed: 32664639
Journal of Neuroinflammation CX3CR1-microglia mediates neuroinflammation and blood pressure regulation in the nucleus tractus solitarii of fructose-induced hypertensive rats Pubmed: 32532282
JOURNAL OF PERIODONTAL RESEARCH Potential biomarkers reflecting inflammation in patients with severe periodontitis: Fractalkine (CX3CL1) and its receptor (CX3CR1) 33641164
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