Anti-CCR4 antibody (ab1669)
- Datasheet
- References (10)
- Protocols
Overview
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Product nameAnti-CCR4 antibody
See all CCR4 primary antibodies -
DescriptionGoat polyclonal to CCR4
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Host speciesGoat
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SpecificityPeptide sequence is < 50 % identical to other human chemokine receptors in this region.
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Tested applicationsSuitable for: Flow Cyt, ICC/IF, ELISA, ICC, IHC-P, WB, IHC-Frmore details
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Species reactivityReacts with: Mouse, Human, Cynomolgus monkey
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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
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Storage bufferPreservative: 0.1% Sodium azide
Constituent: 0.1% BSA -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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.
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| 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
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FunctionHigh 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.
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Tissue specificityPredominantly 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.
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Sequence similaritiesBelongs to the G-protein coupled receptor 1 family.
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Post-translational
modificationsIn natural killer cells, CCL22 binding induces phosphorylation on yet undefined Ser/Thr residues, most probably by beta-adrenergic receptor kinases 1 and 2. -
Cellular localizationCell membrane.
- Information by UniProt
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Database links
- Entrez Gene: 1233 Human
- Entrez Gene: 12773 Mouse
- GenBank: NP_005499.1 Human
- Omim: 604836 Human
- SwissProt: P51679 Human
- SwissProt: P51680 Mouse
- Unigene: 184926 Human
- Unigene: 1337 Mouse
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Alternative names
- C C chemokine receptor type 4 antibody
- C C CKR 4 antibody
- C-C chemokine receptor type 4 antibody
see all
Images
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Flow Cytometry - Anti-CCR4 antibody (ab1669)This image is courtesy of an anonymous Abreviewab1669 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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR4 antibody (ab1669)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.
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Immunohistochemistry (Frozen sections) - Anti-CCR4 antibody (ab1669)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)).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR4 antibody (ab1669)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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR4 antibody (ab1669)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.
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Immunohistochemistry - Anti-CCR4 antibody (ab1669)Immunohistochemistry using ab1669 on a section of human spleen.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR4 antibody (ab1669)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. -
Western blot - Anti-CCR4 antibody (ab1669)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.
Protocols
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) »
