Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Rat, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Neutralization, Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Bioassay, In vivo assay

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2B Clone # 247506
View all CXCR4 Primary Antibodies »

Product Specifications

Immunogen

Y3 rat myeloid cell line transfected with mouse CXCR4
Met1-Ser359
Accession # P70658

Specificity

Detects mouse CXCR4 transfectants. Does not stain irrelevant transfectants.

Clonality

Monoclonal

Host

Rat

Isotype

IgG2B

Endotoxin Level

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

Scientific Data Images for Mouse CXCR4 Antibody

Detection of CXCR4 antibody in Mouse Thymocytes antibody by Flow Cytometry.

Detection of CXCR4 in Mouse Thymocytes by Flow Cytometry.

Mouse thymocytes were stained with (A) Rat Anti-Mouse CXCR4 Monoclonal Antibody (Catalog # MAB21651) or (B) Rat IgG2B isotype control antibody (Catalog # MAB0061) followed by anti-Rat IgG PE-conjugated Secondary Antibody (Catalog # F0105B) and Rat anti-Mouse CD8 alpha APC-conjugated Monoclonal Antibody (Catalog # FAB116A). View our protocol for Staining Membrane-associated Proteins.
CXCR4 antibody in Mouse Neural Progenitor Cells by Immunocytochemistry (ICC).

CXCR4 in Mouse Neural Progenitor Cells.

CXCR4 was detected in immersion fixed mouse neural progenitor cells using 10 µg/mL Rat Anti-Mouse CXCR4 Monoclonal Antibody (Catalog # MAB21651) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
CXCR4 antibody in Mouse Splenocytes by Immunocytochemistry (ICC).

CXCR4 in Mouse Splenocytes.

CXCR4 was detected in immersion fixed mouse splenocytes using 10 µg/mL Rat Anti-Mouse CXCR4 Monoclonal Antibody (Catalog # MAB21651) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
CXCR4 antibody in Mouse Spleen by Immunohistochemistry (IHC-Fr).

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.

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

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

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

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

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

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.

Applications for Mouse 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: Mouse thymocytes

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed mouse neural progenitor cells and immersion fixed mouse splenocytes

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed frozen sections of mouse spleen

Neutralization

Measured by its ability to neutralize CXCL12/SDF‑1 alpha -induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with mouse CXCR4. The Neutralization Dose (ND50) is typically 8‑40 µg/mL in the presence of 10 ng/mL Recombinant Mouse CXCL12/SDF‑1 alpha.

Reviewed Applications

Read 5 reviews rated 4.4 using MAB21651 in the following applications:

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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

UniProt

P70658

Additional CXCR4 Products

Product Documents for Mouse CXCR4 Antibody

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Product Specific Notices for Mouse CXCR4 Antibody

For research use only

Citations for Mouse CXCR4 Antibody

Customer Reviews for Mouse CXCR4 Antibody (5)

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Customer Images

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  • Mouse CXCR4 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: BMM stem cells
    Species: Mouse
    Verified Customer | Posted 08/20/2021
    Mouse CXCR4 Antibody MAB21651
  • Mouse CXCR4 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Hepa 1-6 mouse hepatoma cell line
    Species: Mouse
    Verified Customer | Posted 03/07/2020
    Mouse CXCR4 Antibody MAB21651
  • Mouse CXCR4 Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: 4T1 mouse breast cancer cell line
    Species: Mouse
    Verified Customer | Posted 03/05/2019
    1:75 Citrate buffer antigen retrieval
    Mouse CXCR4 Antibody MAB21651
  • Mouse CXCR4 Antibody
    Name: Travis Durkin
    Application: Block/Neutralize
    Sample Tested: BaF3 mouse pro-B cell line
    Species: Mouse
    Verified Customer | Posted 12/18/2017
    > 95% monomer
    Mouse CXCR4 Antibody MAB21651
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 22752632
    Species: Mouse
    Verified Customer | Posted 02/19/2015

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