Mouse CXCR4 Antibody

CXCR4 in Mouse Spleen.

CXCR4 was detected in immersion fixed frozen sections of mouse spleen using Rat Anti-Mouse CXCR4 Monoclonal Antibody (Catalog # MAB21651) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS017) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

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

Recombinant Mouse CXCL12/SDF-1a (Catalog # 460-SD) chemo-attracts the BaF3 mouse pro-B cell line transfected with mouse 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 Mouse CXCL12/ SDF-1a (10 ng/mL) is neutral-ized (green line) by increasing concentrations of Rat Anti-Mouse CXCR4 Monoclonal Antibody (Catalog # MAB21651). The ND₅₀ is typically 8-40 µg/mL.

Detection of Mouse CXCR4 by Western Blot

Effects of SDF-1-CXCR4/CXCR7 pathway on MSC chemotaxis in vitro.(A) The chemotaxis in response to SDF-1 alpha (10 ng/ml for 12 h) was performed in the NP-MSCs and HP-MSCs treated with a neutralizing anti-CXCR4 antibody, an anti-CXCR7 antibody, and the respective isotype-matched control antibodies. *P<0.05, vs NP-MSCs; †P<0.05, vs the respective isotype-matched control antibodies. (B) NP-MSCs were transiently overexpressed with CXCR4 using pORF9-mCXCR4 vector or with CXCR7 using pORF9-mCXCR7 vector (n = 6). A negative control empty (pORF9-MCS) vector was used. (C) The transfected cells were subjected to chemotaxis in response to the indicated concentrations of SDF-1 alpha for 12 h. *P<0.05, vs the empty vector. (D and E) Western blot analysis (D) and FCM (E) were performed to determine the intracellular and extracellular expression of both CXCR4 and CXCR7 in the cells treated with or without SDF-1 alpha (50 ng/ml for 60 min). (F) The chemotaxis in response to SDF-1 alpha was performed in the cells treated with or without SDF-1 alpha. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/22511954), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse CXCR4 by Immunocytochemistry/Immunofluorescence

Downregulation of Cxcr4, SSEA1 and expression of TRA98 after release from GoST induction.(A) Immunofluorescence of Cxcr4 (white) and SSEA1 (green) after release from GoST induction. Cxcr4 and SSEA1 were detected simultaneously. DNA was counterstained with DAPI (blue). Cxcr4 and SSEA1 were expressed in ES, cES and GoST cells. Cxcr4 was downregulated at 4, 8 and 12 days after release from GoST induction, while SSEA1 remained expressed until 4 days after release from GoST induction and was downregulated afterwards at 8 and 12 days after release. Scalebar = 50 μm. (B) Immunofluorescence of TRA98 (white) and SSEA1 (green) after release from GoST induction. TRA98 and SSEA1 were detected simultaneously. DNA was counterstained with DAPI (blue). TRA98 and SSEA1 were expressed in ES, cES and GoST cells. Expression of TRA98 slightly decreased but remained detectable in the nucleus at 4, 8 and 12 days after release from GoST induction, while SSEA1 remained expressed until 4 days after release from GoST induction and was downregulated afterwards at 8 and 12 days after release. Scalebar = 50 μm. See Fig. S6. Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/srep25104), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CXCR4 by Functional

CXCL12 induces CXCR4/CXCR7 and cell polarization in HUVECs. HUVECs were cultured on plastic Petri dishes (pretreated with PDL) with BSA stripes (the control group) and BSA plus CXCL12 stripes (CXCL12 group) for 5 min. HUVECs were fixed and stained with antibodies against CXCR4 (purple), CXCR7 (blue) and F-Actin (red). The micro stripes of fluorescein-conjugated BSA (A) or CXCL12 (H) are shown. CXCR4 (B), CXCR7 (C) and F-Actin (D) in HUVECs cultured on BSA stripes stayed in the resting state, while the polarization of CXCR4 (arrow: I), CXCR7 (arrow: J) and F-Actin (arrow: K) was observed in HUVECs cultured on CXCL12 stripes for 5 min. Moreover, the HUVECs in the BSA control group showed no morphological changes (E–G). The CXCL12 group HUVECs shows polarization towards the CXCL12 stripe (L–N). Scale bar: 10 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28811579), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CXCR4 by Functional

CXCL12 induces CXCR4/CXCR7 and cell polarization in HUVECs. HUVECs were cultured on plastic Petri dishes (pretreated with PDL) with BSA stripes (the control group) and BSA plus CXCL12 stripes (CXCL12 group) for 5 min. HUVECs were fixed and stained with antibodies against CXCR4 (purple), CXCR7 (blue) and F-Actin (red). The micro stripes of fluorescein-conjugated BSA (A) or CXCL12 (H) are shown. CXCR4 (B), CXCR7 (C) and F-Actin (D) in HUVECs cultured on BSA stripes stayed in the resting state, while the polarization of CXCR4 (arrow: I), CXCR7 (arrow: J) and F-Actin (arrow: K) was observed in HUVECs cultured on CXCL12 stripes for 5 min. Moreover, the HUVECs in the BSA control group showed no morphological changes (E–G). The CXCL12 group HUVECs shows polarization towards the CXCL12 stripe (L–N). Scale bar: 10 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28811579), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse CXCR4 by Immunohistochemistry

RhoA modulates the CXCR4-CXCL12 axis in breast tumors. (a) Representative immunohistochemical (IHC) staining images of breast primary tumor sections of mice orthotopically injected with 4T1 GFP-LUC cells carrying indicated lentiviral modifications, stained with antibodies for CD31 (top, left), F4/80 (middle, left) and alpha-SMA (bottom, left). CD31+ area (top, right), F4/80+ cells (middle, right) and alpha-SMA+ area (bottom, right) per tumor field of view (FOV) (mean ± SE) quantifications of IHC images. N = 4. (b) Representative IHC staining images of breast primary tumor sections of above mice stained with CXCL12 antibody (left). CXCL12+ area per tumor FOV (mean ± SE) quantification of IHC images. N = 4. (c) Representative immunofluorescent images of breast primary tumor sections of above mice co-stained with alpha-SMA, CXCL12 and DRAQ5 nuclear stain, showing the co-localization of alpha-SMA and CXCL12 signals. Scale bar: 50 µm. (d) Representative IHC staining images of breast primary tumor sections of above mice stained with CXCR4 antibody (left). CXCR4+ area per tumor FOV (mean ± SE) quantification of IHC images. N = 4. (e) Relative mCXCR4 mRNA levels normalized with mActin (mean ± SE), as quantified by qRT-PCR in primary breast tumors of above mice (top) and in 4T1 GFP-LUC cells carrying indicated lentiviral modifications (bottom). N = 4, ns- not significant *P < 0.05, unpaired Student’s t-test (two-tailed). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31705019), licensed under a CC-BY license. Not internally tested by R&D Systems.