製品の概要

  • 製品名
  • 製品の詳細
    Goat polyclonal to CCR4
  • 特異性
    Peptide sequence is < 50 % identical to other human chemokine receptors in this region.
  • アプリケーション
    適用あり: Flow Cyt, ICC/IF, ELISA, ICC, IHC-P, WB, IHC-Frmore details
  • 種交差性
    交差種: Mouse, Human, Cynomolgus monkey
  • 免疫原

    Synthetic peptide:

    SNYYLYESIPKPCTKEGIKAFGE

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

  • ポジティブ・コントロール
    • 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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
Flow Cyt Use at an assay dependent concentration.
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.

ターゲット情報

  • 機能
    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.
  • 組織特異性
    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.
  • 配列類似性
    Belongs to the G-protein coupled receptor 1 family.
  • 翻訳後修飾
    In natural killer cells, CCL22 binding induces phosphorylation on yet undefined Ser/Thr residues, most probably by beta-adrenergic receptor kinases 1 and 2.
  • 細胞内局在
    Cell membrane.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

画像

  • 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 minutesCCR4 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

参考文献

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

レビューと Q&A

Merci beaucoup pour toutes ces informations détailléesqui nous sont très utiles pour identifier l'origine du problème rencontré avec le ab1669 lot GR34771-3.

Je n'ai rien à modifier à votre prot...

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Merci de nous avoir contactés.

Nous sommes désolés d'apprendre que le produit ab1669 que vous avez reçu ne fonctionne pas comme attendu.

Nos produits sont garantis pour fonctionner tels qu'ils sont dé...

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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

投稿 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

投稿 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

投稿 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

投稿 Dec 28 2011

Thank you for your enquiry. Ab1667: Antigen Retrieval: Sections were treated with antigen retrieval solution (Dako # S-1700, pH 6.0) and steamed for 20 minutes followed by cooling for 20 minutes. Variations of the method will work successfully as...

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Thank you for your patience. The originator of ab12953 and ab13396 has gotten back to me with the standard IHC protocol that they follow which I have included below. If you have any additional questions, please let me know. Tissue Preparation: ...

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Thank you for your phone call. For ab13268, ab1662, and ab1669, the originator has informed me that the antigen retrieval method for IHC-formalin fixed, paraffin embedded sections involved treating the sections with antigen retrieval solution (Dako # S...

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Thank you for submitting the completed questionnaire to us and for your patience. We are very sorry to hear that you are having problem with this antibody. We have searched our database and found that this is a popular selling product and your feedb...

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1-10 of 11 Abreviews or Q&A

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

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