Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Rabbit

Applications

Validated:

Immunohistochemistry, Neutralization, Flow Cytometry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Immunoprecipitation, Binding Assay, ELISA Detection, Functional Assay

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 44716
View all CXCR4 Primary Antibodies »

Product Specifications

Immunogen

3T3 cells transfected with human CXCR4

Specificity

Detects human CXCR4. It will also react with cells expressing feline CXCR4 but not rat CXCR4.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human CXCR4 Antibody

Detection of CXCR4 antibody in Jurkat Human Cell Line antibody by Flow Cytometry.

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 antibody in Human Lymph Node by Immunohistochemistry (IHC-P).

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 antibody in Human Spleen by Immunohistochemistry (IHC-P).

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.

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

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

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

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-…), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human CXCR4 by Immunohistochemistry

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-…), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Sheep CXCR4 by Immunohistochemistry

Detection of Sheep CXCR4 by Immunohistochemistry

Inflammatory markers. Sections of activated ovine spleen, isolated from a sheep with endocarditis. Hematoxylin and eosin (H&E, top left) staining reveals the characteristic splenic red pulp (R) and white pulp (W), with the central arterioles (CA). Sections are stained with antibodies against CD45, CD3, CD64, EGF-like module-containing mucin-like hormone receptor-like 1 (EMR-1), colony stimulating factor-1 receptor (CSF-1R), interleukin-10 (IL-10), inducible nitric oxide synthase (iNOS), CD163, CD200R, CD44, Arginase-1 (Arg-1), CCR-7, CXCR4, and tumor necrosis factor-alpha (TNF-alpha ). Scale bars, 200 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30159315), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CXCR4 by Western Blot

Detection of CXCR4 by Western Blot

Knockdown of CXCR4 abrogates trastuzumab resistance. CXCR4 was silenced by specific shRNA in HCC1419 cells (“Materials and methods” section). The puromycin-resistant stable colonies were pooled together and named shCXCR4. A pool of cells infected with the lentivirus containing a non-effective vector (shRNA-NE) was selected and used as the control. A Western blot analysis was used to confirm the reduction in CXCR4 expression. B Illustration of co-cultures. C. BCAFs and HMFs were cultured in Dulbecco modified Eagle medium/nutrient mixture/F-12 supplemented with 10% FBS for 72 h. The culture supernatant was collected and tested for SDF-1 alpha using ELISA following the manufacturer’s instructions. The medium used for the cell culture was used as the negative control. The data were analyzed with one-way ANOVA. D, E CXCR4-knockdown cells or non-silent control cells were co-cultured with BCAFs (D) or with BCAFs and PBMCs (E) in 3D, followed by treatment with trastuzumab as illustrated in B. At the endpoint of the study, relative cell viability was quantitatively analyzed using CellTiter-Glo 3D viability assay kit. The data were analyzed with one-way ANOVA (*P < 0.05, **P < 0.01, ***P < 0.001 compared with the non-silent control cells). F, G CXCR4-knockdown cells or non-silent control cells were used for trastuzumab-induced antibody-dependent cellular cytotoxicity (detail in “Materials and methods” section). The cells were stained with propidium iodide and analyzed by flow cytometry (F). Data were analyzed using t-test analysis of variance and are reported as the mean ± SD of triplicates (G) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37280713), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CXCR4 by Western Blot

Detection of CXCR4 by Western Blot

Targeting CXCR4 abrogates trastuzumab resistance. A HER2 + breast cancer cells with or without primary trastuzumab resistance were examined for CXCR4 expression with its antibody (UMB2, Abcam) using western blot analysis. The density of the bands was quantitatively analyzed. B, C Cell lines with high expression of CXCR4 (HCC1419 and HCC202) and low expression of CXCR4 (BT474 and SKBR3) were seeded in 3D Matrigel and treated with trastuzumab (B) or AMD3100 with SDF-1 alpha (4 ng/ml). (C). The total area of the acini was quantitatively analyzed (see “Materials and methods” section). D HCC1419 cells grown in 3D Matrigel culture were treated with trastuzumab (2 µg/ml), AMD3100 (1 µM), SDF-1 alpha (4 ng/ml), or the combination. Photographs were taken on day 13 after the start of treatment. The total area of the acini was quantitatively analyzed using AlphaView SA software (E). F–I Clonogenic assay. HCC1419 (F) and HCC202 (H) cells were seeded at low density and treated with AMD3100 (0.5 µM), SDF-1 alpha (4 ng/ml), trastuzumab (2.5 µg/ml), or the combination. The plates were scanned on day 18 after the start of treatment. Colony formation was quantitatively analyzed using AlphaView SA software. E, G, I Data were analyzed using one-way ANOVA and are reported as mean ± SD of triplicates, representing two independent experiments (*P < 0.0001 compared with vehicle) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37280713), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CXCR4 by Western Blot

Detection of CXCR4 by Western Blot

Knockdown of CXCR4 abrogates trastuzumab resistance. CXCR4 was silenced by specific shRNA in HCC1419 cells (“Materials and methods” section). The puromycin-resistant stable colonies were pooled together and named shCXCR4. A pool of cells infected with the lentivirus containing a non-effective vector (shRNA-NE) was selected and used as the control. A Western blot analysis was used to confirm the reduction in CXCR4 expression. B Illustration of co-cultures. C. BCAFs and HMFs were cultured in Dulbecco modified Eagle medium/nutrient mixture/F-12 supplemented with 10% FBS for 72 h. The culture supernatant was collected and tested for SDF-1 alpha using ELISA following the manufacturer’s instructions. The medium used for the cell culture was used as the negative control. The data were analyzed with one-way ANOVA. D, E CXCR4-knockdown cells or non-silent control cells were co-cultured with BCAFs (D) or with BCAFs and PBMCs (E) in 3D, followed by treatment with trastuzumab as illustrated in B. At the endpoint of the study, relative cell viability was quantitatively analyzed using CellTiter-Glo 3D viability assay kit. The data were analyzed with one-way ANOVA (*P < 0.05, **P < 0.01, ***P < 0.001 compared with the non-silent control cells). F, G CXCR4-knockdown cells or non-silent control cells were used for trastuzumab-induced antibody-dependent cellular cytotoxicity (detail in “Materials and methods” section). The cells were stained with propidium iodide and analyzed by flow cytometry (F). Data were analyzed using t-test analysis of variance and are reported as the mean ± SD of triplicates (G) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37280713), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CXCR4 by Western Blot

Detection of CXCR4 by Western Blot

Targeting CXCR4 abrogates trastuzumab resistance. A HER2 + breast cancer cells with or without primary trastuzumab resistance were examined for CXCR4 expression with its antibody (UMB2, Abcam) using western blot analysis. The density of the bands was quantitatively analyzed. B, C Cell lines with high expression of CXCR4 (HCC1419 and HCC202) and low expression of CXCR4 (BT474 and SKBR3) were seeded in 3D Matrigel and treated with trastuzumab (B) or AMD3100 with SDF-1 alpha (4 ng/ml). (C). The total area of the acini was quantitatively analyzed (see “Materials and methods” section). D HCC1419 cells grown in 3D Matrigel culture were treated with trastuzumab (2 µg/ml), AMD3100 (1 µM), SDF-1 alpha (4 ng/ml), or the combination. Photographs were taken on day 13 after the start of treatment. The total area of the acini was quantitatively analyzed using AlphaView SA software (E). F–I Clonogenic assay. HCC1419 (F) and HCC202 (H) cells were seeded at low density and treated with AMD3100 (0.5 µM), SDF-1 alpha (4 ng/ml), trastuzumab (2.5 µg/ml), or the combination. The plates were scanned on day 18 after the start of treatment. Colony formation was quantitatively analyzed using AlphaView SA software. E, G, I Data were analyzed using one-way ANOVA and are reported as mean ± SD of triplicates, representing two independent experiments (*P < 0.0001 compared with vehicle) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37280713), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human CXCR4 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: Jurkat human acute T cell leukemia cell line

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human lymph node and human spleen

Neutralization

Measured by its ability to neutralize CXCL12/SDF‑1 alpha -induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CXCR4. The Neutralization Dose (ND50) is typically 2.5-12 µg/mL in the presence of 1 ng/mL Recombinant Human/Feline/Rhesus Macaque CXCL12/SDF‑1 alpha.

Reviewed Applications

Read 6 reviews rated 5 using MAB172 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CXCR4

CXCR4, also known as CD184, is a G-protein-linked seven transmembrane spanning receptor that binds stromal cell-derived factor-1 (SDF-1). CXCR4 acts as a co-factor for T-cell tropic HIV-1 and -2 viral entry into cells. While primarily a membrane protein, CXCR4 undergoes trafficking and internalization in response to stimulation with phorbol esters and ligand (1). Cytoplasmic and nuclear localization of CXCR4 has been observed in colorectal and renal carcinomas (2,3) and it has been used as the basis of prognosis and metastatic state (3,4,5).

References

  1. Orsini, M.J. et al. (1999) J. Biol. Chem. 274:31076.
  2. Zagzag, D. et al. (2005) Cancer Res. 65:6178.
  3. Speetjens, F.M. et al. (2009) Cancer Microenvironment 2:1.
  4. Wang, L. et al. (2009) Oncology Reports 22:1333.
  5. Amara, S. et al. (2015) Cancer Biomark. 15:869.

Long Name

C-X-C Motif Chemokine Receptor 4

Alternate Names

CD184, D2S201E, FB22, Fusin, HM89, HSY3RR, LAP-3, LAP3, LCR1, LESTR, NPY3R, NPYRL, NPYY3R, WHIMS

Entrez Gene IDs

7852 (Human); 12767 (Mouse); 60628 (Rat); 483900 (Canine); 493676 (Feline)

Gene Symbol

CXCR4

Additional CXCR4 Products

Product Documents for Human CXCR4 Antibody

Certificate of Analysis

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Product Specific Notices for Human CXCR4 Antibody

For research use only

Citations for Human CXCR4 Antibody

Customer Reviews for Human CXCR4 Antibody (6)

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Customer Images

Showing  1 - 5 of 6 reviews Showing All
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  • Human CXCR4 Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Tumor tissue and Ductal carcinoma
    Species: Human
    Verified Customer | Posted 08/03/2021
    Human CXCR4 Antibody MAB172
  • Human CXCR4 Antibody
    Name: Anonymous
    Application: Microarray
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 12/07/2020
    Antibody was printed on custom arrays and incubated with fluorescently labeled human EDTA plasma
    Human CXCR4 Antibody MAB172
  • Human CXCR4 Antibody
    Name: Anonymous
    Sample Tested: SUDHL-6 and Ramos human Burkitt's lymphoma cell line
    Species: Human
    Verified Customer | Posted 11/16/2018
    Cell line stained with human CXCR4 for 45 min and secondary antibody anti-mouse FITC for 45min &amp; Isotype control and unstained cell used in experiment
    Human CXCR4 Antibody MAB172
  • Human CXCR4 Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: SUDHL-6 and Ramos cells and SU-DHL-6 cell line
    Species: Human
    Verified Customer | Posted 10/27/2018
    Cell line stained with human CXCR4 for 45 min and secondary antibody anti-mouse FITC for 45min (Isotype control and unstained cell used in experiment)
    Human CXCR4 Antibody MAB172
  • Human CXCR4 Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Human cell line
    Species: Human
    Verified Customer | Posted 03/15/2018
    Human CXCR4 Antibody MAB172
  • Human CXCR4 Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Jurkat
    Species: Human
    Verified Customer | Posted 03/09/2018
    Human CXCR4 Antibody MAB172

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Protocols

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