Overview

  • Product name
  • Description
    Goat polyclonal to CCR4
  • Host species
    Goat
  • Specificity
    Peptide sequence is < 50 % identical to other human chemokine receptors in this region.
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, ELISA, ICC, IHC-P, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Human, Cynomolgus monkey
  • Immunogen

    Synthetic peptide:

    SNYYLYESIPKPCTKEGIKAFGE

    , corresponding to amino acids 17-39 of Human CCR4 (extracellular domain).

  • Positive control
    • Human peripheral blood cells or paraffin sections of spleen. IHC-P:FFPE mouse lymph node normal. IHC-P:FFPE human tonsil normal. IHC-P:FFPE rat spleen normal.

Properties

Applications

Our Abpromise guarantee covers the use of ab1669 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.

ab37373 - Goat polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration. Fix cells with 4% PFA (see Ritter et al).
ELISA 1/100000.
ICC 1/300.
IHC-P 1/300.
WB 1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 42 kDa).
IHC-Fr Use at an assay dependent concentration. Fix in acetone at -20°C for 5 minutes. See Heller et al.

Target

  • Function
    High affinity receptor for the C-C type chemokines CCL17/TARC and CCL22/MDC. The activity of this receptor is mediated by G(i) proteins which activate a phosphatidylinositol-calcium second messenger system. Can function as a chemoattractant homing receptor on circulating memory lymphocytes and as a coreceptor for some primary HIV-2 isolates. In the CNS, could mediate hippocampal-neuron survival.
  • Tissue specificity
    Predominantly expressed in the thymus, in peripheral blood leukocytes, including T-cells, mostly CD4+ cells, and basophils, and in platelets; at lower levels, in the spleen and in monocytes. Detected also in macrophages, IL-2-activated natural killer cells and skin-homing memory T-cells, mostly the ones expressing the cutaneous lymphocyte antigen (CLA). Expressed in brain microvascular and coronary artery endothelial cells.
  • Sequence similarities
    Belongs to the G-protein coupled receptor 1 family.
  • Post-translational
    modifications
    In natural killer cells, CCL22 binding induces phosphorylation on yet undefined Ser/Thr residues, most probably by beta-adrenergic receptor kinases 1 and 2.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • C C chemokine receptor type 4 antibody
    • C C CKR 4 antibody
    • C-C chemokine receptor type 4 antibody
    • C-C CKR-4 antibody
    • CC CKR 4 antibody
    • CC-CKR-4 antibody
    • CCR 4 antibody
    • CCR-4 antibody
    • CCR4 antibody
    • CCR4_HUMAN antibody
    • CD194 antibody
    • Chemokine (CC motif) receptor 4 antibody
    • chemokine C C motif receptor 4 antibody
    • ChemR13 antibody
    • CKR4 antibody
    • CMKBR 4 antibody
    • CMKBR4 antibody
    • HGCN 14099 antibody
    • K5 5 antibody
    • K5-5 antibody
    • MGC88293 antibody
    see all

Images

  • ab1669 staining CCR4 in Mouse whole blood by Flow Cytometry. Red blood cells were lysed in PBS + 1% BSA and 0.01% sodium azide and fixed in paraformaldehyde. The sample was incubated with the primary antibody (1/100 in PBS + 1% BSA and 0.01% sodium azide) for 1 hour at 4°C. A biotin-conjugated Donkey anti-goat IgG polyclonal (1/200) was used as the secondary antibody.
    Gating Strategy: Live Lymphocytes.

    See Abreview

  • IHC image of CD3 staining in a formalin fixed, paraffin embedded normal rat spleen tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab1669 at 1/100 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    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.

     

  • Frozen sections of  human skin tissue stained for CCR4 using ab1669 in immunohistochemical analysis (green). CD31(red) and nuclei (blue) are also shown. The right hand panel is a merged image.

    Image courtesy of PMID 25915746 (PLoS One 2015 Apr 27;10(4)).

  • IHC image of CD3 staining in a formalin fixed, paraffin embedded normal human tonsil tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab1669 at 1/100 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    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.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • IHC image of CD3 staining in TISSUE formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab1669 at 1/100 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    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.

  • Immunohistochemistry using ab1669 on a section of human spleen.

  • IHC image of CCR4 staining in human spleen formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab1669, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. 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.
  • Anti-CCR4 antibody (ab1669) at 1 µg/ml + Human thymus tissue lysate - total protein (ab30146) at 10 µg

    Secondary
    Rabbit polyclonal to Goat IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 55 kDa (why is the actual band size different from the predicted?)
    Additional bands at: 100 kDa, 37 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 12 minutes


    CCR4 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

References

This product has been referenced in:
  • Maolake A  et al. Tumor-associated macrophages promote prostate cancer migration through activation of the CCL22-CCR4 axis. Oncotarget 8:9739-9751 (2017). Human . Read more (PubMed: 28039457) »
  • Berlato C  et al. A CCR4 antagonist reverses the tumor-promoting microenvironment of renal cancer. J Clin Invest 127:801-813 (2017). Read more (PubMed: 28134623) »

See all 10 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

Application
Flow Cytometry
Sample
Cynomolgus Monkey Cell (Whole blood)
Specification
Whole blood
Preparation
Cell harvesting/tissue preparation method: After red blood cell lysis
Sample buffer: PBS/BSA 1%/Azide 0.01%
Fixation
Paraformaldehyde
Permeabilization
No
Gating Strategy
alive lymphocytes
Username

Abcam user community

Verified customer

Submitted Apr 27 2012

Abcam has not validated the combination of species/application used in this Abreview.
Application
Flow Cytometry
Sample
Rat Cell (Splenocytes)
Specification
Splenocytes
Preparation
Cell harvesting/tissue preparation method: spleen was mashed
Sample buffer: PBS/BSA 1%/Azide 0.01%
Permeabilization
No
Gating Strategy
alive lymphocytes
Username

Abcam user community

Verified customer

Submitted Apr 18 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Mouse Cell (Whole blood)
Specification
Whole blood
Preparation
Cell harvesting/tissue preparation method: After red blood cell lysis
Sample buffer: PBS/BSA 1%/Azide 0.01%
Fixation
Paraformaldehyde
Permeabilization
No
Gating Strategy
alive lymphocytes
Username

Abcam user community

Verified customer

Submitted Apr 18 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (PBMC)
Specification
PBMC
Preparation
Cell harvesting/tissue preparation method: after ficoll
Sample buffer: PBS/BSA 1%/ Azide 0.01%
Permeabilization
No
Gating Strategy
on monocytes
Username

Abcam user community

Verified customer

Submitted Dec 28 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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