Overview

  • Product nameAnti-CXCR4 antibody [UMB2]
    See all CXCR4 primary antibodies
  • Description
    Rabbit monoclonal [UMB2] to CXCR4
  • Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human CXCR4 aa 300 to the C-terminus (C terminal).
    (Peptide available as ab155072)

  • Positive control
    • Human cervical carcinoma, ovarian adenocarcinoma and tonsil tissues IF/ICC: Jurkat cells. IHC-P: FFPE Brain of Mouse E14 embryo
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Alternative versions available:

    Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203) Knockout validated

    Anti-CXCR4 antibody (Alexa Fluor® 647) [UMB2] (ab208129)

     

     

Applications

Our Abpromise guarantee covers the use of ab124824 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 1/100. Predicted molecular weight: 39 kDa.Can be blocked with CXCR4 peptide (ab155072).
IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

ICC/IF Use a concentration of 5 µg/ml.

Target

  • FunctionReceptor 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 specificityExpressed 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 diseaseDefects 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 similaritiesBelongs to the G-protein coupled receptor 1 family.
  • DomainThe 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 localizationCell 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

Anti-CXCR4 antibody [UMB2] images

  • All lanes : Anti-CXCR4 antibody [UMB2] (ab124824)

    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
    Goat anti-rabbit at 1/10000 dilution

    Predicted band size : 39 kDa
    Observed band size : 41 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 ab124824 (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 ab124824 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.

  • Immunofluorescence staining of Jurkat cells with purified ab124824 at a working dilution of 1 in 250, counter-stained with DAPI. Tubulin was stained with mouse anti-tubulin at a dilution of 1/1000 (ab7291) and Alexa Fluor® 594 goat anti-mouse at a dilution of 1/500 (ab150120) . The secondary antibody was ab150077 Alexa Fluor® 488 goat anti rabbit, used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in the bottom middle and right hand panels - for the first negative control, purified ab124824 was used at a dilution of 1/200 followed by an Alexa Fluor® 555 goat anti-mouse antibody at a dilution of 1/500 and for the second negative control mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab15007) were used.

  • Anti-CXCR4 antibody [UMB2] (ab124824) at 1/100 dilution (purified) + Rat muscle tissue lysate at 20 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Anti-CXCR4 antibody [UMB2] (ab124824) at 1/100 dilution (purified) + NIH/3T3 cell lysate at 20 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Anti-CXCR4 antibody [UMB2] (ab124824) at 1/100 dilution (purified) + WI-38 cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • ab124824 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 (ab124824 at 5ug/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.

  • Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab124824 at a working dilution of 1/500. The secondary antibody used is ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Unpurified ab124824, at 1/50 dilution, staining CXCR4 in paraffin-embedded Human cervical carcinoma tissue by immunohistochemistry.

  • Unpurified ab124824, at 1/50 dilution, staining CXCR4 in paraffin-embedded Human ovarian adenocarcinoma tissue by immunohistochemistry.

  • Unpurified ab124824, at 1/50 dilution, staining CXCR4 in paraffin-embedded Human tonsil tissue by immunohistochemistry.

  • Lane 1 : Anti-CXCR4 antibody [UMB2] (ab124824) at 1/500 dilution (unpurified)
    Lane 2 : Anti-CXCR4 antibody [UMB2] (ab124824) at 1/500 dilution

    Lane 1 : HEK239 tansfected with a CXCR4 (mouse) expression vector cell lysate
    Lane 2 : HEK239 tansfected with an empty expression vector cell lysate

    Lysates/proteins at 100000 cells per lane.

    Secondary
    HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 39 kDa
    Observed band size : 42,47 kDa (why is the actual band size different from the predicted?)


    Exposure time : 10 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview

References for Anti-CXCR4 antibody [UMB2] (ab124824)

This product has been referenced in:
  • Wang Y  et al. Polyplex-mediated inhibition of chemokine receptor CXCR4 and chromatin-remodeling enzyme NCOA3 impedes pancreatic cancer progression and metastasis. Biomaterials 101:108-20 (2016). WB . Read more (PubMed: 27267632) »
  • Zhang L  et al. CXCR4 activation promotes differentiation of human embryonic stem cells to neural stem cells. Neuroscience N/A:N/A (2016). WB ; Human . Read more (PubMed: 27615032) »

See all 30 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Monkey Cell (Kidney)
Permeabilization Yes - 0.1% triton x-100
Specification Kidney
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative Paraformaldehyde
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Submitted Jun 10 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Brain)
Permeabilization No
Specification Brain
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative Paraformaldehyde
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Submitted Mar 04 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Heart)
Permeabilization Yes - 0.2% triton TX-100
Specification Heart
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative Formaldehyde
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Submitted Feb 24 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (BEL-7404)
Gel Running Conditions Non-reduced Denaturing (10%)
Loading amount 40 µg
Treatment 80µM Doxorubicin for 24hrs
Specification BEL-7404
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. guoqing zhu

Verified customer

Submitted Jan 20 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Heart)
Gel Running Conditions Reduced Denaturing (10% Invitrogen gel)
Loading amount 40 µg
Specification Heart
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Mr. Dan Bromage

Verified customer

Submitted Dec 30 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (1BR3.G fibrblast)
Gel Running Conditions Reduced Denaturing (10%)
Loading amount 50 µg
Specification 1BR3.G fibrblast
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5%
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Submitted Jul 22 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (1BR3.G fibroblast)
Permeabilization Yes - Triton 1%
Specification 1BR3.G fibroblast
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: rt°C
Fixative Formaldehyde
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Submitted Jul 22 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (colon)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: EDTA, pH8, 100C, 20 min
Permeabilization No
Specification colon
Blocking step 3% H2O2 as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative Formaldehyde
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Submitted Oct 27 2014

Application Immunohistochemistry (Frozen sections)
Blocking step 0.3% Triton X-100, 10mg/mL BSA, 5% Normal Goat Serum, in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Sample Dog Tissue sections (Atria)
Specification Atria
Permeabilization Yes - 0.3% Triton X-100, 10mg/mL BSA, 5% Normal Goat Serum, in PBS
Fixative Paraformaldehyde
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Dr. Juan Estrada

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Submitted Jul 28 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - other (HEK239)
Gel Running Conditions Reduced Denaturing
Loading amount 100000 cells
Treatment transfected with cxcr4 (mouse) plasmid or transfected with the empty expression vector
Specification HEK239
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Nov 19 2013

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