Application Note
Chemokine C-X3-C-Motif Ligand 1 (CX3CL1) also known as 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. 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. Thus, chemotaxis assay used 24-well microchemotaxis system was undertaken to detect the chemotactic effect of CX3CL1 on the human monocytic cell line THP-1. Briefly, THP-1 cells were seeded into the upper chambers (150 μl cell suspension, 1x10⁶ cells/ml in RPMI-1640 with FBS free) and SLC (15. 625 ng/ml, 31. 25 ng/ml, 62. 5 ng/ml and 125 ng/ml diluted separately in serum free RPMI-1640) was added in lower chamber with a polycarbonate filter (8um pore size) used to separate the two compartments. After incubation at 37ºC with 5% CO₂ for 1h, the filter was removed, then cells in low chamber were observed by inverted microscope at low magnification (×100) and the number of migrated cells were counted at high magnification (×400) randomly. And CX3CL1 is able to induce migration of THP-1 cells.
Observed MW
37 kDa.
Form
Lyophilized powder
Buffer
Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
Preservative
ProClin 300
Storage
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
Region/Sequence
N-terminal His-Tag; Gln83~Pro328 (NP_604450.1)
Expression System
E. coli
Purity
> 95%
Endotoxin
< 1 EU/μg
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
Synonyms
C-X3-C motif chemokine ligand 1 , Cx3c , Scyd1
Background
chemokine that binds its receptor CX3CR1; induces chemotaxis and increased intracellular calcium levels in microglia [RGD, Feb 2006]
Database
Research Area