Human CXCR4 Antibody

Chemotaxis Induced by CXCL12/SDF‑1 alpha and Neutralization by Human CXCR4 Antibody.

Recombinant Human/Feline/Rhesus Macaque CXCL12/SDF-1a (Catalog # 350-NS) chemoattracts the BaF3 mouse pro-B cell line transfected with human CXCR4 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/Feline/Rhesus Macaque CXCL12/SDF-1a (1 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human CXCR4 Monoclonal Antibody (Catalog # MAB173). The ND₅₀ is typically 1-5 µg/mL.

Detection of Canine CXCR4 by Immunocytochemistry/Immunofluorescence

Ectopic expression of CXCR7 and CXCR4 on MDCK cells.(A) MDCK were stably transfected with empty vector (Mock), with CXCR7, CXCR4, a vector coding for a CXCR7 lacking the cytoplasmic C-terminus ( delta CXCR7), and a vector coding for a chimeric CXCR7 containing the DRYLAIV motive of CXCR4. Receptor expression was determined by FACS analysis using saturating antibody concentrations (see Methods). (B) Confocal immunofluorescence analysis of unfixed MDCK cells expressing CXCR7 (upper panels) or CXCR4 (lower panels). Cells also expressed N-ter-Lck mCherry as membrane marker (red fluorescence). Left panels: confocal images of planes cut through intracellular regions of MDCK monolayers. Right panels: x-z planes reconstructed from confocal x-y stacks. For receptor (green) detection anti-CXCR7 (11G8 R&D) or anti CXCR4 (MAB173 R&D) were used. Receptor-bound primary antibodies were revealed with goat anti mouse IgG conjugated with Alexa488 (green fluorescence). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/20161793), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of CXCR4 by Western Blot

In vitro cell invasion is stimulated by the SDF-1/CXCR4 pathway independently from HPV status.The modulation of E6 expression was monitored using Western-blot in HPV-positive HeLa (A) and TC-1 (B) cells and in HPV-negative B16F10 (C) cells, after 3 and 6 days of incubation with Cidofovir (CDF). Modulation of P53 expression was assessed in HeLa cells after CDF incubation (A). Cell invasion was measured using a Matrigel assay in HeLa (A), TC-1 (B) and B16F10 (C) cells. Recombinant human CXCL12/SDF-1 (100 ng/mL; R&D Systems) was used as chemoattractant and modulation of cell migration was recorded after treatment with CXCR4-blocking antibody or/and Cidofovir (CDF). The invasion rate was determined by counting crystal violet-stained cells. Invasion was stimulated by SDF-1 alpha /CXCR4 independently from the HPV status of the cells but Cidofovir anti-invasive action was restricted to the two HPV-positive cell lines. Three independent experiments with three chambers each time were performed. ***P<0.01, for a statistically true difference, as compared to the untreated group. # P<0.01 compared to SDF-1 alpha -treated group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/19325708), licensed under a CC-BY license. Not internally tested by R&D Systems.