Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-CXCR4 antibody [EPUMBR3] - Low endotoxin, Azide free (ab222223)

Overview

  • Product name

    Anti-CXCR4 antibody [EPUMBR3] - Low endotoxin, Azide free
    See all CXCR4 primary antibodies
  • Description

    Rabbit monoclonal [EPUMBR3] to CXCR4 - Low endotoxin, Azide free
  • Host species

    Rabbit
  • Specificity

    This antibody recognizes only the non-phosphorylated C-terminus of CXCR4 (residues 341-352). Phosphorylation of S346/347 blocks antibody binding. PMID: 24154522, 25451233.

    We recommend dephosphorylation of samples using lambda phosphatase treatment. Please refer to application notes.

  • Tested applications

    Suitable for: Flow Cyt, 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. FC: Jurkat cells.
  • General notes

    ab222223 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab222223 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
Flow Cyt Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 39 kDa.

We recommend lambda protein phosphatase treatment of the membrane prior to primary antibody incubation (PMID 24154522). Use 800U for 1 hr at RT then rinse in wash buffer three times.

IHC-P Use at an assay dependent concentration.

We recommend lambda protein phosphatase treatment prior to IHC processing (PMID 24154522). Use 800U for 1 hr at RT then rinse in PBS three times.

ICC/IF Use at an assay dependent concentration.
  • 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

    • Flow Cytometry analysis of Jurkat (human T cell leukemia T lymphocyte) cells labeling CXCR4 with purified ab181020 at 1:200 dilution (10.23 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - GreenThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab222223)
    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181020).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181020).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181020).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181020).

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181020).

    References

    ab222223 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    There are currently no Customer reviews or Questions for ab222223.
    Please use the links above to contact us or submit feedback about this product.

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    Sign up