Anti-CXCR4 (extracellular) Antibody

C-X-C chemokine receptor type 4, CD184, Fusin, LESTR, NPYRL, SDF-1 receptor, WHIMS
    Cat #: ACR-014
    Alternative Name C-X-C chemokine receptor type 4, CD184, Fusin, LESTR, NPYRL, SDF-1 receptor, WHIMS
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
    Immunogen
    • Peptide (C)EGISIYTSDNYTEE, corresponding to amino acid residues 2-15 of human CXCR4 (Accession P61073). Extracellular, N-terminus.
    • Anti-CXCR4 (extracellular) Antibody
    Accession (Uniprot) Number P61073
    Gene ID 7852
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - 12/14 amino acid residues identical; rat - 9/14 amino acid residues identical.
    RRID AB_10613475.
    Purity Affinity purified on immobilized antigen.
    Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Isotype Rabbit IgG.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 25 μl, 50 μl, or 0.2 ml double distilled water (DDW), depending on the sample size.
    Antibody concentration after reconstitution 0.8 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
    Negative control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
    Negative control antigen reconstitution 100 µl double distilled water (DDW).
    Negative control antigen storage after reconstitution -20°C.
    Preadsorption Control 1 μg peptide per 1 μg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, if, ifc, ih, wb
    May also work in: ip*, lci*
    Western blot
    • Anti-CXCR4 (extracellular) Antibody
      Western blot analysis of rat (lanes 1 and 3) and mouse (lanes 2 and 4) brain membranes:
      1,2. Anti-CXCR4 (extracellular) Antibody (#ACR-014), (1:200).
      3,4. Anti-CXCR4 (extracellular) Antibody, preincubated with the negative control antigen.
    Immunohistochemistry
    • Anti-CXCR4 (extracellular) Antibody
      Expression of CXCR4 in rat cerebellum
      Immunohistochemical staining of rat cerebellum frozen section using Anti-CXCR4 (extracellular) Antibody (#ACR-014), (1:100). CXCR4 is expressed in Purkinje cell bodies and axonal prolongations (arrows). Hoechst 33342 is used as the counterstain.
    • Anti-CXCR4 (extracellular) Antibody
      Expression of CXCR4 in rat brain
      Immunohistochemical staining of rat temporal cortex using Anti-CXCR4 (extracellular) Antibody (#ACR-014), (1:400) followed by donkey anti-rabbit-biotin and streptavidin labeled with Cy3. A. CXCR4 staining (red) appears in neuronal soma and processes (arrows). DAPI is used as the counterstain (blue).
    Immunocytochemistry
    • Human vascular smooth muscle cells (hVSMCs) (Gao, X. et al. (2018) Sci. Rep. 8, 2730.).
    Indirect flow cytometry
    • Anti-CXCR4 (extracellular) Antibody
      Cell surface detection of CXCR4 in Jurkat living cells:
      ___ Unstained cells.
      ___ Cells + Anti-CXCR4 (extracellular) Antibody (#ACR-014), (10 µg/0.5x106 cells).
    • The negative control antigen is not suitable for this application.
    References
    1. Raman, D. et al. (2007) Cancer Lett. 256, 137
    2. Burger, J.A. and Kipps, T.J. (2006) Blood 107, 1761.
    3. Loetscher, P. et al. (2000) Adv. Immunol. 74, 127.
    4. Feng, Y. et al. (1996) Science 272, 872.
    5. Wong, D. and Korz, W. (2008) Clin. Cancer. Res. 14, 7975.
    6. Vila-Coro, A.J. et al. (1999) FASEB J. 13, 1699.
    7. Holland, J.D. et al. (2006) Cancer Res. 66, 4117.
    8. Richardson, R.M. et al. (2003) J. Biol. Chem. 278, 15867.
    9. Zou, Y.R. et al. (1998) Nature 393, 595.
    10. Heesen, M. et al. (1996) J. Immunol. 157, 5455.
    11. Habasque, C. et al. (2002) Mol. Hum. Reprod. 8, 419.
    Scientific background

    Chemokines are small molecular weight, soluble secreted proteins that bind and activate their respective G-protein coupled receptor (GPCR), chemokine receptors in order to evoke a cellular response resulting in migration or chemotaxis1.

    The chemokine system involves more than 40 chemokines and 18 chemokine receptors. The receptors are designated CXCR1-5, CCR1-11, XCR1 and CX3CR1, based on their specific ligand preference2.

    Chemokine receptors are present on many different cell types. They were initially detected on leukocytes, where they were found to play an important role in the migration of these cells to inflammation sites3.

    CXCR4 was originally identified as an orphan receptor, and soon gained much attention when it was discovered as a coreceptor for HIV-14. Besides from being involved in HIV-1 infection/progression, CXCR4 is found to be upregulated in many different cancers/tumors and has evolved to become a target for the development of antagonists5. CXCL12 (SDF-1α) is the sole ligand for CXCR4. Following binding of its ligand, CXCR4 undergoes dimerization and activates Gi G-proteins6,7. However downstream activation through CXCR4 could also occur through other G-proteins and non-G-proteins5. The down regulation of the CXCR4 receptor is initiated by phosphorylation of its cytoplasmic tail, which is followed by the binding of arrestin. The receptor is then internalized through endocytosis and degraded in the lysosome. Downregulation of CXCR4 could also occur through the stimulation of other GPCRs8.

    The distribution of CXCR4 is quite broad and involves the central nervous system (CNS)9, spleen10, testes11, hematopoietic and non-hematopoietic cells2.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

    Species reactivity key:

    H- Human, M- Mouse, R- Rat
    Last update: 29/07/2020

    Anti-CXCR4 (extracellular) Antibody (#ACR-014) is a highly specific antibody directed against an extracellular epitope of the human CXCR4 chemokine receptor. The antibody can be used for western blot, immunocytochemistry, immunohistochemistry, and indirect flow cytometry applications. It has been designed to recognize CXCR4 from rat, mouse and human samples.

    For research purposes only, not for human use

    Applications

    Specifications

    Scientific Background

    Citations

    Citations
    Immunocytochemistry citations
    1. Human vascular smooth muscle cells (hVSMCs).
      Gao, X. et al. (2018) Sci. Rep. 8, 2730.
    2. Human vascular smooth muscle cells (hVSMCs).
      Albee, L.J. et al. (2017) J. Am. Heart Assoc. 6, e006575.
    Indirect flow cytometry citations
    1. Human vascular smooth muscle cells (hVSMCs).
      Gao, X. et al. (2018) PLoS ONE 13, e0204041.
    2. Rat mesenchymal stem cells.
      Li, N. et al. (2015) Am. J. Transl. Res. 7, 1058.
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