ELISA Kit for Chemokine C-X3-C-Motif Ligand 1 (CX3CL1)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-104 93
EDTA plasma(n=5) 81-93 85
heparin plasma(n=5) 80-90 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) 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) 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) 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) 86-101% 91-101% 78-88% 79-97%
EDTA plasma(n=5) 85-99% 94-101% 95-102% 94-105%
heparin plasma(n=5) 78-99% 97-105% 88-103% 83-92%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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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
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