Human CXCR4 Antibody

Detection of CXCR4 in Jurkat Human Cell Line by Flow Cytometry. Jurkat human acute T cell leukemia cell line was stained with Mouse Anti-Human CXCR4 Monoclonal Antibody (Catalog # MAB172, filled histogram) or isotype control antibody (Catalog # MAB004, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.

CXCR4 in Human Lymph Node. CXCR4 was detected in immersion fixed paraffin-embedded sections of human lymph node using 15 µg/mL Human CXCR4 Monoclonal Antibody (Catalog # MAB172) overnight at 4 °C. Tissue was stained with the Anti-Mouse HRP-AEC Cell & Tissue Staining Kit (red; Catalog # CTS003) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

CXCR4 in Human Spleen. CXCR4 was detected in immersion fixed paraffin-embedded sections of human spleen using Mouse Anti-Human CXCR4 Monoclonal Antibody (Catalog # MAB172) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

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 # MAB172). The ND₅₀ is typically 2.5-12 µg/mL.

Detection of Human CXCR4 by Flow Cytometry CXCR4 neutralization leads to attenuation of the CD44+/CD133+ prostate progenitor population.(A) DU145 cells were plated in 96-well low-attachment plates at 100 cells per well (5 replicates) and the spheres were grown in serum-free, EBM medium with supplements. The antibodies were replenished daily. Cells were imaged with an Acumen eX3 microplate cytometer and spheres were detected using image analysis software. The sphere size was measured by GFP intensity. The spheres were discriminated from cell debris using a Gaussian filter. The spheres included in the analysis are outlined in red and indicated by arrows. Representative data from one of two independent experiments is shown; *- p value<0.05. (B) Flow cytometry analysis revealed attenuation of CD44+/CD133+ population in DU145 cells treated with 10 µg/ml neutralizing anti-CXCR4 (mouse monoclonal IgG, clone 44716, R&D Systems) or 10 µg/ml control antibody (mouse IgG isotype control, Lifespan Bioscience Inc.) for 5 days. The cell were grown in medium supplemented with 2% FBS. Culture medium was refreshed every second day; *- p value<0.05. (C) Western blot analysis of DU145 cells treated with 10 µg/ml neutralizing anti-CXCR4 antibody for 5 days demonstrated downregulation of the PI3K/AKT pathway activation compared to the cells treated with 10 µg/ml control antibody. The cell were grown in medium supplemented with 2% FBS. Culture medium was refreshed every second day. (D) Preincubation of prostate cancer cells with neutralizing anti-CXCR4 antibody significantly delays tumor growth. 5×105 DU145 cells pretreated with neutralizing anti-CXCR4 or control antibody for 5 days were embedded in BD matrigel and injected s.c. into NOD/SCID mice.*- p value<0.01. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0031226), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CXCR4 by Immunohistochemistry CXCR4 and CD133 are highly expressed in ovarian cancer patients. (A) Representative panel of immunohistochemistry for CD133 and CXCR4 expression (Magnification 200X; insert 400X). Non-homogeneous and focal cytoplasmic and membranous CD133 expression rated as negative (I), focally low (1–10% cancer cells) (II), focally high CD133 expression (>10% stained cancer cells) (III). Extensive homogeneous cytoplasmic and membranous, CXCR4 staining was reported rated as negative (IV) moderate (<50% of cancer cells) (V) and high expression (>50% of cancer cells) (VI). (B) QPCR analysis of CXCR4 and CD133 in freshly resected ovarian tumors and corresponding normal ovarian tissue. Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/srep10357), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CXCR4 by Immunohistochemistry Immunohistochemical staining for MIF and CXCR4 in human esophageal carcinoma. Our data showed low expression levels of MIF (a) and CXCR4 (c) (X 400) and high expression levels of MIF (b) and CXCR4 (d), compared with the negative control (e) (X 400), in tumor tissues from patients with ESCC. The arrows point to the positive staining of tumor cells or TILs. Image collected and cropped by CiteAb from the following publication (https://translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-60), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CXCR4 by Immunohistochemistry Immunohistochemical staining for MIF and CXCR4 in human esophageal carcinoma. Our data showed low expression levels of MIF (a) and CXCR4 (c) (X 400) and high expression levels of MIF (b) and CXCR4 (d), compared with the negative control (e) (X 400), in tumor tissues from patients with ESCC. The arrows point to the positive staining of tumor cells or TILs. Image collected and cropped by CiteAb from the following publication (https://translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-60), licensed under a CC-BY license. Not internally tested by R&D Systems.