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    products/primary-antibodies/cxcr4-antibody-umb2-ab124824.pdf

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Immunology Innate Immunity Chemokines Alpha Chemokine Rec. (CXCR)
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RecombinantRabMAb

Recombinant Anti-CXCR4 antibody [UMB2] (ab124824)

  • Datasheet
  • SDS
Reviews (17)Q&A (6)References (188)

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Flow Cytometry (Intracellular) - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Immunohistochemistry (Frozen sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Western blot - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Immunohistochemistry (Frozen sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Western blot - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Western blot - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Western blot - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Western blot - Anti-CXCR4 antibody [UMB2] (ab124824)
  • Anti-CXCR4 antibody [UMB2] (ab124824)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [UMB2] to CXCR4
  • Suitable for: IHC-Fr, Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free

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Overview

  • Product name

    Anti-CXCR4 antibody [UMB2]
    See all CXCR4 primary antibodies
  • Description

    Rabbit monoclonal [UMB2] to CXCR4
  • Host species

    Rabbit
  • Specificity

    Although some customers can get this ab to work in mouse and rat successfully we cannot reproduce this in house in IHC so cannot guarantee it. We would recommend antibody Anti-CXCR4 antibody [EPUMBR3] (ab181020) for use in mouse IHC.

    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: IHC-Fr, Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human CXCR4 aa 300 to the C-terminus (C terminal). The exact sequence is proprietary.
    (Peptide available as ab155072)

  • Positive control

    • WB: HeLa, Jurkat and WI-38 cell lysates; HEK239 transfected with CXCR4, cell lysate.IF/ICC: Jurkat cells. IHC-P: Human cervical carcinoma, bladder cancer tissue, ovarian adenocarcinoma tissue and tonsil tissue. Flow Cyt (intra): Jurkat cells IHC-Fr: Mouse and rat E14.5 cerebrum
  • General notes

    Our internal data indicates that mouse and rat are not recommended for IHC.

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    UMB2
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Chemokines
    • Alpha Chemokine Rec. (CXCR)
    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Neuroscience
    • Neurology process
    • Growth and Development
    • Axonal Guidance Proteins
    • Stem Cells
    • Neural Stem Cells
    • Surface Molecules
    • Stem Cells
    • Hematopoietic Progenitors
    • Surface Molecules
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells
    • Stem Cells
    • Endothelial Progenitors
    • Endothelial Markers
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Immunology
    • Immune System Diseases
    • Antiviral Signaling
    • HIV-related
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Types of disease
    • Cancer
    • Neuroscience
    • Development

Associated products

  • Alternative Versions

    • Anti-CXCR4 antibody [UMB2] - BSA and Azide free (ab197203)
    • Alexa Fluor® 488 Anti-CXCR4 antibody [UMB2] (ab208128)
    • Alexa Fluor® 647 Anti-CXCR4 antibody [UMB2] (ab208129)
    • Anti-p130 (phospho T986) antibody [EPR2389(2)] (ab211928)
    • Alexa Fluor® 555 Anti-CXCR4 antibody [UMB2] (ab211982)
    • Alexa Fluor® 594 Anti-CXCR4 antibody [UMB2] (ab211984)
    • Anti-p130 (phospho T986) antibody [EPR2389(2)] - BSA and Azide free (ab251534)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human CXCR4 protein (ab155716)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • Anti-CD8 alpha antibody [YTS169.4] (ab22378)
    • Anti-CD34 antibody [MEC 14.7] (ab8158)

Applications

The Abpromise guarantee

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
IHC-Fr (3)
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB (9)
1/100. Predicted molecular weight: 39 kDa.

Being CXCR4 a membrane protein, we recommend to not boil the samples after the lysis (before loading the samples on the WB gel).

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 (2)
1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

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 (3)
Use a concentration of 5 µg/ml.
Notes
IHC-Fr
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
1/100. Predicted molecular weight: 39 kDa.

Being CXCR4 a membrane protein, we recommend to not boil the samples after the lysis (before loading the samples on the WB gel).

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
1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

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 a concentration of 5 µg/ml.

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.
  • Target information above from: UniProt accession P61073 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7852 Human
    • Entrez Gene: 12767 Mouse
    • Entrez Gene: 60628 Rat
    • Omim: 162643 Human
    • SwissProt: P61073 Human
    • SwissProt: P70658 Mouse
    • SwissProt: O08565 Rat
    • Unigene: 593413 Human
    • Unigene: 1401 Mouse
    • Unigene: 44431 Rat
    see all
  • 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 (Intracellular) - Anti-CXCR4 antibody [UMB2] (ab124824)
    Flow Cytometry (Intracellular) - Anti-CXCR4 antibody [UMB2] (ab124824)

    Intracellular Flow Cytometry analysis of Jurkat (human T cell leukemia T lymphocyte) cells labeling CXCR4 with purified ab124824 at 1/260 dilution (10 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 - Green

     

  • Immunohistochemistry (Frozen sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
    Immunohistochemistry (Frozen sections) - Anti-CXCR4 antibody [UMB2] (ab124824)

    Immunohistochemical (Frozen) analysis of mouse E14.5 cerebrum labeling CXCR4 with ab124824 at 1/250 (10.8 μg/mL). Sections were fixed with 4% PFA and permeabilised with 0.2% Triton X-100. ab150077 lexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/mL) was used as the secondary antibody. Nuclei were counterstained blue with DAPI. Heat mediated antigen retrieval was performed using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). 

    Positive staining on mouse embryonic cerebrum.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)Costa, M.J. et al PLoS One. 2018 Mar 19;13(3):e0194688. doi: 10.1371/journal.pone.0194688. eCollection 2018 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Immunohistochemical detection of CXCR4 expression in human tissue specimens of normal appearance

    CXCR4 was detected in the indicated PFA-fixed, paraffin-embedded human tissues using ab124824 at 5 μ/ml overnight at 4°C.

    A, kidney. B, adrenal gland. C, cerebellum. D, bone marrow, brown staining: CXCR4, green staining: CD45. E, Spleen. F, testis. G, lung. H, colon.

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

  • Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] (ab124824)
    Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] (ab124824)Fischer, T. et al PLoS One. 2008;3(12):e4069. doi: 10.1371/journal.pone.0004069. Epub 2008 Dec 31 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Characterization of UMB-2 (ab124824) by immunofluorescent staining of transfected cells. HEK-293 cells expressing CCR7 or CXCR4 were either not exposed or exposed to 100 ng/ml MIP-3 or 100 ng/ml SDF-1 for 30 min, subsequently fixed and immunofluorescently stained with 1 µg/ml anti-CCR7 {1188} or anti-CXCR4 {UMB-2} at a dilution of 1∶100. Note that UMB-2 detected prominent immunofluorescence at the level of the plasma membrane only in CXCR4- but not in CCR7-expressing cells, and that SDF-1 exposure induced a rapid translocation of CXCR4 receptor immunostaining from the plasma membrane into the cytosol. Representative results from one of three independent experiments are shown. Scale bar, 20 µm.

  • Immunohistochemistry (Frozen sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
    Immunohistochemistry (Frozen sections) - Anti-CXCR4 antibody [UMB2] (ab124824)

    Immunohistochemical (Frozen) analysis of rat E14.5 cerebrum labeling CXCR4 with ab124824 at 1/250 (10.8 μg/mL). Sections were fixed with 4% PFA and permeabilised with 0.2% Triton X-100. ab150077 lexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/mL) was used as the secondary antibody. Nuclei were counterstained blue with DAPI. Heat mediated antigen retrieval was performed using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). 

    Positive staining on rat embryonic cerebrum.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)Li X et al., European journal of nuclear medicine and molecular imaging. 45,4 (2018): 558-566. Fig 3. doi:10.1007/s00259-017-3831-0. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Immunohistochemistry analysis of cryosections from carotid endarterectomy specimens labeling CXCR4 with ab124824 at 1/300 dilution. CXCR4 expression in a representative inflamed carotid plaque lesion. Brightfield micrographs showed brown chemoimmunoreactive CXCR4 and CD68 (macrophage) staining. Co-localized CXCR4 and CD68 expression was observed in these two adjacent sections.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)

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

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] (ab124824)
    Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] (ab124824)

    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.

  • Western blot - Anti-CXCR4 antibody [UMB2] (ab124824)
    Western blot - Anti-CXCR4 antibody [UMB2] (ab124824)Fischer, T. et al PLoS One. 2008;3(12):e4069. doi: 10.1371/journal.pone.0004069. Epub 2008 Dec 31. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

    Western blot analysis of the specificity of anti-CXCR4 antibodies. Membrane preparations from HEK-293 cells stably transfected to express either CCR7 or CXCR4 were separated on 10% SDS-polyacrylamide gels and blotted onto nitrocellulose membranes. Membranes were then incubated with affinity-purified 1 µg/ml anti-CCR7 {1188} or anti-CXCR4 {UMB-2} hybridoma supernatant at a dilution of 1:100. Blots were developed using enhanced chemiluminescence. Note that UMB-2 detected a band of the expected molecular weight only in CXCR4- but not in CCR7-transfected cells. Two additional experiments gave similar results.

  • Western blot - Anti-CXCR4 antibody [UMB2] (ab124824)
    Western blot - Anti-CXCR4 antibody [UMB2] (ab124824)
    All lanes : Anti-CXCR4 antibody [UMB2] (ab124824) at 1/1000 dilution

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lanes 2-3 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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


    Exposure time: 1 second


    Blocking and diluting buffer: 5% NFDM/TBST

    We suggest to not boil the sample after lysis.

  • Western blot - Anti-CXCR4 antibody [UMB2] (ab124824)
    Western blot - Anti-CXCR4 antibody [UMB2] (ab124824)
    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)

    Immunohistochemical staining of paraffin embedded human bladder cancer 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] (ab124824)
    Immunocytochemistry/ Immunofluorescence - Anti-CXCR4 antibody [UMB2] (ab124824)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)

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

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCR4 antibody [UMB2] (ab124824)

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

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-CXCR4 antibody [UMB2] (ab124824)
    Western blot - Anti-CXCR4 antibody [UMB2] (ab124824)This image is courtesy of an anonymous Abreview
    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 transfected with a CXCR4 (mouse) expression vector cell lysate
    Lane 2 : HEK239 transfected with an empty expression vector cell lysate

    Lysates/proteins at 100000 cells per lane.

    Secondary
    All lanes : 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

    See Abreview

  • Anti-CXCR4 antibody [UMB2] (ab124824)
    Anti-CXCR4 antibody [UMB2] (ab124824)

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (188)

Publishing research using ab124824? Please let us know so that we can cite the reference in this datasheet.

ab124824 has been referenced in 188 publications.

  • Chen S  et al. Exosomes derived from retinoblastoma cells enhance tumour deterioration by infiltrating the microenvironment. Oncol Rep 45:278-290 (2021). PubMed: 33416154
  • Feng W  et al. CXCL12-mediated HOXB5 overexpression facilitates Colorectal Cancer metastasis through transactivating CXCR4 and ITGB3. Theranostics 11:2612-2633 (2021). PubMed: 33456563
  • Liang Q  et al. Stromal cell-derived factor-1/Exendin-4 cotherapy facilitates the proliferation, migration and osteogenic differentiation of human periodontal ligament stem cells in vitro and promotes periodontal bone regeneration in vivo. Cell Prolif 54:e12997 (2021). PubMed: 33511708
  • Dimachkie MD  et al. Exploration of biomarkers from a pilot weight management study for men undergoing radical prostatectomy. Urol Oncol 39:495.e7-495.e15 (2021). PubMed: 33563536
  • Si H  et al. Anti-Tumor Effect of Celastrol on Hepatocellular Carcinoma by the circ_SLIT3/miR-223-3p/CXCR4 Axis. Cancer Manag Res 13:1099-1111 (2021). PubMed: 33574707
View all Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-CXCR4 antibody [UMB2]

Average
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 04 2016

Immunohistochemistry (Frozen sections) abreview for Anti-CXCR4 antibody [UMB2]

Good
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 24 2016

Immunohistochemistry (Frozen sections) abreview for Anti-CXCR4 antibody [UMB2]

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Juan Estrada

Verified customer

Submitted Jul 28 2014

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