Human Fractalkine ELISA Kit

Human Fractalkine ELISA Kit Features

Product Name           Human Fractalkine ELISA Kit
Assay Type                   Solid Phase Sandwich ELISA
Format                           96-well strip plate
Assay Length              4.5 hours
Sample Volume         Serum ,Plasma, cell culture media , 100ul
Detection Method    Colorimetric
Sensitivity                   0.2 ng/mL
Assay Range                0.25-60 ng/mL
Reactivity                     Human
Cross-Reactivity       No cross-reactivity observed with available related molecules
Gene Names               CX3CL1;  ABCD-3, C3Xkine, CXC3, CXC3C, NTN, NTT, SCYD1, fractalkine, neurotactin
Gene ID                         6376
Swissprot ID               P78423

Recovery

Linearity

 Standard Curve: Human Fractalkine ELISA Kit

Kit Components

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent
• Standards
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Protocol Outline

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 µl of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4°C.
4. Wash and add 100 µl of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Wash and add 100 µl of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Wash and add 100 µl of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 µl of Stop Solution to each well.
11. Read at 450 nm/570 nm.

Background

Fractalkine is a large cytokine protein of 373 amino acids, it contains multiple domains and is the only known member of the CX3C chemokine family. Its gene is located on human chromosome 16along with some CC chemokines known as CCL17 and CCL22. CX3CL1 is produced as a long protein with an extended mucin-like stalk and a chemokine domain on top. The mucin-like stalk permits it to bind to the surface of certain cells.

Soluble CX3CL1 potently chemoattracts T cells and monocytes, while the cell-bound chemokine promotes strong adhesion of leukocytes to activated endothelial cells, where it is primarily expressed. CX3CL1 elicits its adhesive and migratory functions by interacting with the chemokine receptor CX3CR1.

Fractalkine is found commonly throughout the brain, particularly in neural cells, and its receptor is known to be present on microglial cells. It has also been found to be essential for microglial cell migration. CX3CL1 is also up-regulated in the hippocampus during a brief temporal window following spatial learning, the purpose of which may be to regulate glutamate-mediated neurotransmission tone. This indicates a possible role for the chemokine in the protective plasticity process of synaptic scaling.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Regulatory Status      

Human Fractalkine ELISA Kit is for Research Use Only.

• Manual
• Assay procedure
• MSDS