Human CXCR4 Antibody (44708)

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 Human CXCR4 Monoclonal Antibody (Catalog # MAB171). The ND₅₀ is typically 0.2-1.2 µg/mL.

Detection of Mouse CXCR4 by Flow Cytometry

Anti-CXCR4 antibody reduces CXCR4 surface expression, and blocks migration toward CXCL12 and BMSCs.(A) Relative MFIR of CXCR4 surface staining after incubation of Raji transfectants with 100 µg/mL of anti-CXCR4 antibody for 30 minutes. (B) Flow cytometry histograms showing the reduction in CXCR4 expression after incubation with neutralizing anti-CXCR4 antibody. White histograms represent negative unstained controls. (C) CXCR4-blocked cells were stimulated with 100 ng/mL of CXCL12 for 5 minutes and activation of Akt and ERK1/2 proteins analyzed by western blotting. (D) CXCR4-blocked cells were subjected to migration assay for 4 hours at 37°C in 5% CO2 toward 100 ng/mL of CXCL12 (left panel) or the stromal MS-5 cell line (right panel). Cells in the lower chamber were counted by flow cytometry under a defined flow rate for 5 minutes. Number of migrated cells treated with isotypic control was considered 100%. Results are shown as the mean ± SEM of 4 independent experiments. (E) Cell viability was measured by Annexin V-PI staining in cells incubated with 100 µg/mL of anti-CXCR4 or isotypic control for 48 hours. Results are shown as the mean ± SEM of 4 independent experiments Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0081221), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse CXCR4 by Western Blot

Anti-CXCR4 antibody reduces CXCR4 surface expression, and blocks migration toward CXCL12 and BMSCs.(A) Relative MFIR of CXCR4 surface staining after incubation of Raji transfectants with 100 µg/mL of anti-CXCR4 antibody for 30 minutes. (B) Flow cytometry histograms showing the reduction in CXCR4 expression after incubation with neutralizing anti-CXCR4 antibody. White histograms represent negative unstained controls. (C) CXCR4-blocked cells were stimulated with 100 ng/mL of CXCL12 for 5 minutes and activation of Akt and ERK1/2 proteins analyzed by western blotting. (D) CXCR4-blocked cells were subjected to migration assay for 4 hours at 37°C in 5% CO2 toward 100 ng/mL of CXCL12 (left panel) or the stromal MS-5 cell line (right panel). Cells in the lower chamber were counted by flow cytometry under a defined flow rate for 5 minutes. Number of migrated cells treated with isotypic control was considered 100%. Results are shown as the mean ± SEM of 4 independent experiments. (E) Cell viability was measured by Annexin V-PI staining in cells incubated with 100 µg/mL of anti-CXCR4 or isotypic control for 48 hours. Results are shown as the mean ± SEM of 4 independent experiments Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0081221), licensed under a CC-BY license. Not internally tested by R&D Systems.