製品の概要

  • 製品名
    Anti-CXCR4 antibody [UMB2] - BSA and Azide free
    CXCR4 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [UMB2] to CXCR4 - BSA and Azide free
  • 由来種
    Rabbit
  • 特異性
    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
  • アプリケーション
    適用あり: Flow Cyt, IHC-P, ICC/IF, IHC-Fr, WBmore details
  • 種交差性
    交差種: Human
    交差が予測される動物種: Mouse, Rat
  • 免疫原

    Synthetic peptide within Human CXCR4 (C terminal). The exact sequence is proprietary.
    (Peptide available as ab155072)

  • ポジティブ・コントロール
    • 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. FC: Jurkat cells
  • 特記事項

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

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

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab197203 in the following tested applications.

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

アプリケーション Abreviews 特記事項
Flow Cyt Use at an assay dependent concentration.
IHC-P 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocol.

ICC/IF Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration. PubMed: 28932900
WB Use at an assay dependent concentration. Predicted molecular weight: 39 kDa.Can be blocked with CXCR4 peptide (ab155072).

Please check the parent abID, ab124824, for more information on dilution ranges.

ターゲット情報

  • 機能
    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.
  • 組織特異性
    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.
  • 関連疾患
    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.
  • 配列類似性
    Belongs to the G-protein coupled receptor 1 family.
  • ドメイン
    The amino-terminus is critical for ligand binding. Residues in all four extracellular regions contribute to HIV-1 coreceptor activity.
  • 翻訳後修飾
    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.
  • 細胞内局在
    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
  • 参照データベース
  • 別名
    • 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

画像

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124824).

  • 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 - GreenThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab197203)
  • 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.

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

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

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124824).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124824).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124824).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124824).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling CXCR4 with purified ab197203 at 1/1000 dilution (1.067 μg/ml). Heat mediated antigen retrieval was performed using Tris/EDTA Buffer, PH9 (ab93684). Hematoxylin was used to counter stain. Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used.

    Membranous with weak cytoplasmic staining on human tonsil.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human B-cell non-hodgkin lymphoma tissue sections labeling CXCR4 with purified ab197203 at 1/1000 dilution (1.067 μg/ml). Heat mediated antigen retrieval was performed using Tris/EDTA Buffer, PH9 (ab93684). Hematoxylin was used to counter stain. Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used.

    Cytoplasmic and membranous staining on tumor cells of human B-cell non-Hodgkin lymphoma.

参考文献

This product has been referenced in:
  • Zhang XQ  et al. Increased protein expression levels of pCREB, BDNF and SDF-1/CXCR4 in the hippocampus may be associated with enhanced neurogenesis induced by environmental enrichment. Mol Med Rep 14:2231-7 (2016). Read more (PubMed: 27432087) »
  • Satomura H  et al. Can expression of CXCL12 and CXCR4 be used to predict survival of gastric cancer patients? Anticancer Res 34:4051-7 (2014). Read more (PubMed: 25075029) »
See all 14 Publications for this product

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