Overview

  • Product name
    Anti-CXCR4 antibody [EPUMBR3]
    See all CXCR4 primary antibodies
  • Description
    Rabbit monoclonal [EPUMBR3] to CXCR4
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human CXCR4. The exact sequence is proprietary.
    Database link: P61073

  • Positive control
    • Human CXCR4 stably expressed in HEK293 cells. Human small cell lung carcinoma tissue. IF/ICC: Jurkat and Ramos cells. IHC-P: FFPE retina of mouse E14 embryo. IHC-P:FFPE brain of mouse E14 embryo.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab181020 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/10000. Predicted molecular weight: 39 kDa.
IHC-P Use at an assay dependent concentration.
ICC/IF 1/500.

For unpurified use at 5 µg/mL.

  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      Receptor for the C-X-C chemokine CXCL12/SDF-1 that transduces a signal by increasing intracellular calcium ions levels and enhancing MAPK1/MAPK3 activation. Acts as a receptor for extracellular ubiquitin; leading to enhance intracellular calcium ions and reduce cellular cAMP levels. Involved in haematopoiesis and in cardiac ventricular septum formation. Plays also an essential role in vascularization of the gastrointestinal tract, probably by regulating vascular branching and/or remodeling processes in endothelial cells. Could be involved in cerebellar development. In the CNS, could mediate hippocampal-neuron survival. Acts as a coreceptor (CD4 being the primary receptor) for HIV-1 X4 isolates and as a primary receptor for some HIV-2 isolates. Promotes Env-mediated fusion of the virus.
    • Tissue specificity
      Expressed in numerous tissues, such as peripheral blood leukocytes, spleen, thymus, spinal cord, heart, placenta, lung, liver, skeletal muscle, kidney, pancreas, cerebellum, cerebral cortex and medulla (in microglia as well as in astrocytes), brain microvascular, coronary artery and umbilical cord endothelial cells. Isoform 1 is predominant in all tissues tested.
    • Involvement in disease
      Defects in CXCR4 are a cause of WHIM syndrome (WHIM) [MIM:193670]; also known as warts, hypogammaglobulinemia, infections and myelokathexis. WHIM syndrome is an immunodeficiency disease characterized by neutropenia, hypogammaglobulinemia and extensive human papillomavirus (HPV) infection. Despite the peripheral neutropenia, bone marrow aspirates from affected individuals contain abundant mature myeloid cells, a condition termed myelokathexis.
    • Sequence similarities
      Belongs to the G-protein coupled receptor 1 family.
    • Domain
      The amino-terminus is critical for ligand binding. Residues in all four extracellular regions contribute to HIV-1 coreceptor activity.
    • Post-translational
      modifications
      Phosphorylated on agonist stimulation. Rapidly phosphorylated on serine and threonine residues in the C-terminal. Phosphorylation at Ser-324 and Ser-325 leads to recruitment of ITCH, ubiquitination and protein degradation.
      Ubiquitinated by ITCH at the cell membrane on agonist stimulation. The ubiquitin-dependent mechanism, endosomal sorting complex required for transport (ESCRT), then targets CXCR4 for lysosomal degradation. This process is dependent also on prior Ser-/Thr-phosphorylation in the C-terminal of CXCR4. Also binding of ARRB1 to STAM negatively regulates CXCR4 sorting to lysosomes though modulating ubiquitination of SFR5S.
      Sulfation on Tyr-21 is required for efficient binding of CXCL12/SDF-1alpha and promotes its dimerization.
      O- and N-glycosylated. Asn-11 is the principal site of N-glycosylation. There appears to be very little or no glycosylation on Asn-176. N-glycosylation masks coreceptor function in both X4 and R5 laboratory-adapted and primary HIV-1 strains through inhibiting interaction with their Env glycoproteins. The O-glycosylation chondroitin sulfate attachment does not affect interaction with CXCL12/SDF-1alpha nor its coreceptor activity.
    • Cellular localization
      Cell membrane. In unstimulated cells, diffuse pattern on plasma membrane. On agonist stimulation, colocalizes with ITCH at the plasma membrane where it becomes ubiquitinated.
    • Information by UniProt
    • Database links
    • Alternative names
      • C-X-C chemokine receptor type 4 antibody
      • CD184 antibody
      • CD184 antigen antibody
      • Chemokine (C X C motif) receptor 4 antibody
      • Chemokine CXC Motif Receptor 4 antibody
      • CXC-R4 antibody
      • CXCR-4 antibody
      • CXCR4 antibody
      • CXCR4_HUMAN antibody
      • D2S201E antibody
      • FB22 antibody
      • Fusin antibody
      • HM89 antibody
      • HSY3RR antibody
      • LAP 3 antibody
      • LAP3 antibody
      • LCR1 antibody
      • LESTR antibody
      • Leukocyte derived seven transmembrane domain receptor antibody
      • Leukocyte-derived seven transmembrane domain receptor antibody
      • Lipopolysaccharide associated protein 3 antibody
      • Neuropeptide Y receptor Y3 antibody
      • NPY3R antibody
      • NPYR antibody
      • NPYRL antibody
      • NPYY3 antibody
      • NPYY3R antibody
      • Probable G protein coupled receptor lcr1 homolog antibody
      • SDF 1 receptor antibody
      • SDF-1 receptor antibody
      • SEVEN-TRANSMEMBRANE-SEGMENT RECEPTOR antibody
      • Stromal cell derived factor 1 receptor antibody
      • Stromal cell-derived factor 1 receptor antibody
      • WHIM antibody
      • WHIMS antibody
      see all

    Images

    • Immunocytochemistry/Immunofluorescence analysis of Ramos (Human Burkitt's lymphoma cell line) labeling CXCR4 with purified ab181020 at 1/500 dilution. Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

    • IHC image of CXCR4 staining on retina of E14 mouse embryo formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with 10 mM sodium citrate buffer (pH6) for 20 mins in a microwave at 600W, and incubated overnight at + 4oC with ab181020 at 5 ugml. Staining of primary antibody was detected using the appropriate biotinylated secondary antibodies followed by incubation with avidin-biotinylated peroxidase solution. DAB was used as the chromogen (15 min). The section was then counterstained with haematoxylin. As a negative control (inset), an identical assay was performed on retina of E14 knockout mouse (CXCR4 -/-) embryo.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    • ab181020 stained Jurkat cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab181020 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

    • All lanes : Anti-CXCR4 antibody [EPUMBR3] (ab181020)

      Lane 1 : CHO (negative control)
      Lane 2 : Jurkat whole cell
      Lane 3 : Jurkat membrane
      Lane 4 : Jurkat nuclear (negative control)

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit at 1/10000 dilution

      Predicted band size: 39 kDa
      Observed band size: 43 kDa (why is the actual band size different from the predicted?)



      Running buffer: MOPS.

      Conditions: denatured/reduced.

      This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with ab181020 (anti-CXCR4) and ab7671 (loading ctrl), overnight at 4°C. Before imaging, antibody binding was detected using labelled goat anti-rabbit (H+L; green) and labelled goat anti-mouse (H+L; red) at 1:10,000 dilutions for 1hr at room temperature.

    • IHC image of CXCR4 staining on Brain of E14 mouse embryo formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with 10 mM sodium citrate buffer (pH6) for 20 mins in a microwave at 600W, and incubated overnight at + 4oC with ab181020 at 1/500 dilution. Staining of primary antibody was detected using the appropriate biotinylated secondary antibodies followed by incubation with avidin-biotinylated peroxidase solution. DAB was used as the chromogen (15 min). The section was then counterstained with haematoxylin. As a negative control (inset), an identical assay was performed on Brain of E14 knockout mouse (CXCR4 -/-) embryo.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    • Anti-CXCR4 antibody [EPUMBR3] (ab181020) at 1/1000 dilution + CXCR4 stably expressed in HEK293 cells

      Predicted band size: 39 kDa

    • Immunohistochemical analysis of paraffin embedded Human small cell lung carcinoma tissue labeling CXCR4 using ab181020.

    References

    This product has been referenced in:

    See 1 Publication for this product

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