Human CX3CL1/Fractalkine Chemokine Domain Antibody

Chemotaxis Induced by CX3CL1/Fractalkine and Neutralization by Human CX3CL1/Fractalkine Antibody.

Recombinant Human CX3CL1/Fractalkine (Catalog # 365-FR) chemoattracts the BaF3 mouse pro-B cell line transfected with mouse CX3CR1 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Human CX3CL1/Fractalkine (100 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human CX3CL1/Fractalkine Chemokine Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF365). The ND₅₀ is typically 0.75-4.5 µg/mL.

Detection of CX3CL1/Fractalkine by Western Blot

CX3CR1 is required for keratinocytes differentiation. (A) Expression of CX3CR1 in parental PA-1 cell line&that stably transfected with either CX3CR1 or scrambled shRNA constructs was examined with WB. Actin served as a loading control. CX3CR1 expression levels quantified using digital densitometry&normalized to the levels of actin expression. (B) Expression of cytokeratin 14 in PA-1 cells stably transfected with either CX3CR1 or scrambled shRNA constructs treated with recombinant BMP-4 or vehicle, as indicated, was examined with immunocytochemistry. CK14 in PA-1 was probed with anti-CK14&Alexa555-conjugated anti-rabbit antibodies,&DNA was stained with 4’,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI); CK14 – red, DNA – blue. Images taken using 5× magnification on the objective. The histogram demonstrates the average intensity of CK14 signal across the field as determined by the Zeiss AxioVision software. Data is an average of three independent experiments. *p < 0.05. (C) Expression of cytokeratins 14&18 in PA-1 cells stably transfected with either CX3CR1 or scrambled shRNA constructs treated with recombinant BMP-4 or vehicle, as indicated, was examined with WB. beta-Tubulin served as a loading control. The histogram shows CK14&CK18 expression levels. Expression of CK18 in CX3CR1shRNA-transfected PA-1 cells was arbitrarily set as 1&expression of both CK14&CK18 in other conditions was calculated accordingly. The data represent a typical WB image,&quantitative analysis was performed using three independently performed experiments. *p < 0.05. (D) Expression of CX3CL1 in SKOV-3&PA-1 cell lines was examined with WB. Actin served as a loading control. SKOV-3 cell line was used as a positive control. Image collected&cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23958497), licensed under a CC-BY license. Not internally tested by R&D Systems.