Antibody Panel to C-C Chemokine Receptor Family - FocusOn 007

CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCR10, CCR11/CCRL1 antibody

Introduction

Chemokines and their receptors play a pivotal role in lymphocyte trafficking, recruiting and recirculation. They act through seven transmembrane G protein-coupled receptors. The chemokine system is increasingly recognized to play an important role in lymphocyte biology. Chemokine receptors are G-protein-coupled receptors (GPCRs) with seven transmembrane (TM7) spanning a-helices belonging to the rhodopsin superfamily. They have an N-terminus outside the cell, three extracellular domains, three intracellular loops and a C-terminus in the cytoplasm wich contains serine and threonine phosphorylation sides. These receptors bind their corresponding ligands with high affinity. Unusualy for members of the TM7 receptor superfamily nearly all chemokine receptors have multiple high-affinity ligands for a single receptor. CCR5 for example binds CCL3 and CCL4 as well as CCL5. Binding of a chemokine to its specific receptor on the cell surface activates a G-protein coupled signal transduction cascade. Finally the activation of protein kinases results in chemotaxis towards the source of the chemokine. Based on their ligand specificity, chemokine receptors can be divided into two major groups, CXCR and CCR, based on the two major classes of chemokines. The C-C Chemokine receptor highlighted in this article were assigned to the cluster of differentiation (CD) group of cell surface molecules during the 8th HLDA workshop in Adelaide 2004 as CD191 (CCR1) to CDw199 (CCR9).

CCR1 was the first CC chemokine receptor identified and binds multiple inflammatory/inducible (see inducible gene) CC chemokines (including CCL4, CCL5, CCL6, CCL14, CCL15, CCL16 and CCL23). In humans, this receptor can be found on peripheral blood lymphocytes and monocytes. There is some suggestion that this chemokine receptor is restricted to memory T-cells within the lymphocyte pool.

Fig. 1: <span>Immunohistochemical staining of spleen lymphocytes using anti- CCR1 antibody SP4001P</span><span>.</span>
Fig. 1: Immunohistochemical staining of spleen lymphocytes using anti- CCR1 antibody SP4001P.
 

CCR2 can interact with CCL2, CCL8 and CCL16 and has been identified on the surface of monocytes, activated memory T cells, B cells, and basophils in humans, and also in peritoneal macrophages in mice.

Fig. 2: <span>Immunohistochemical staining of Spleen (Lymphocytes) using anti- CCR2 antibody SP4086P</span><span>.</span>
Fig. 2: Immunohistochemical staining of Spleen (Lymphocytes) using anti- CCR2 antibody Cat.-No. SP4086P.
 

CCR3 is a receptor for multiple inflammatory/inducible CC chemokines, including CCL11, CCL26, CCL7, CCL13, CCL15, CCL24 and CCL5 that attract eosinophils, and CCL28 that attracts B and T lymphocytes to mucosal tissues. It is most highly expressed in both eosinophils and basophils, but can also be found in Th1 and Th2 cells and airway epithelial cells.

Fig. 3: <span>Immunohistochemical staining of nasal mucosa (allergic rhinitis) using anti- CCR3 antibody SP4002P.</span>
Fig. 3: Immunohistochemical staining of nasal mucosa (allergic rhinitis) using anti- CCR3 antibody Cat.-No. SP4002P.
 

CCR4 is expressed on Th2 T lymphocytes and is up-regulated by T cell receptor activation. However, some reports suggest a role for this receptor also in trafficking of dendritic cells. The CC chemokines CCL3, CCL5, CCL17 and CCL22 signal through this receptor.

Fig. 4: <span>Spectral Confocal Microscopy of CHO cells using KH-4F5 antibody (Cat.#DM1232). CHO cells were transiently transfected with an expression vector encoding CCR4. Binding of KH-4F5 was visualized with a FITC-conjugated secondary antibody (green). Actin filaments are labeled with Alexa Fluor-555 Phalloidin (red). Cell nuclei are stained with DAPI (blue).</span>
Fig. 4: Spectral Confocal Microscopy of CHO cells using KH-4F5 antibody (Cat.-No. DM1232). CHO cells were transiently transfected with an expression vector encoding CCR4. Binding of KH-4F5 was visualized with a FITC-conjugated secondary antibody (green). Actin filaments are labeled with Alexa Fluor-555 Phalloidin (red). Cell nuclei are stained with DAPI (blue). 
 

CCR5 is expressed on several cell types including peripheral blood-derived dendritic cells, CD34+ hematopoietic progenitor cells and certain activated/memory Th1 lymphocytes. This receptor is well defined as a major coreceptor implicated in susceptibility to HIV-1 infection and disease. This receptor has several CC chemokine ligands including CCL2, CCL3, CCL4, CCL5, CCL11, CCL13, CCL14 and CCL16.

Fig. 5: <span>Immunohistochemical analysis of paraffin embedded rectal carcinoma sections, staining CCR5 in cytoplasm, DAB chromogenic reaction.</span>
Fig. 5: Immunohistochemical analysis of paraffin embedded rectal carcinoma sections, staining CCR5 in cytoplasm, DAB chromogenic reaction. Cat.-No. AP23286PU-N.
 

CCR6, a receptor for CCL20, is expressed on unactivated memory T-cells and some dendritic cells. CCR6 is also expressed on Th17 cells.

Fig. 6: <span>CCR6 polyclonal antibody</span><span>.</span>
Fig. 6: CCR6 polyclonal antibody. Cat.-No. AP23322PU-N.
 

CCR7 is a highly important receptor with a role in trafficking of B and T lymphocytes and dendritic cells to and across high endothelial venules and positioning those cells correctly in T cell zones of secondary lymphoid organs. Its ligands include the related chemokines CCL19 and CCL21.

Fig. 7: <span>AP15408PU CCR7 antibody staining of Human Small Intestine</span><span>.</span>
Fig. 7: AP15408PU CCR7 antibody staining of Human Small Intestine.
 

CCR8 is associated with Th2 lymphocytes and is therefore found predominantly in the thymus (in humans) although some expression can be found in the brain, spleen, lymph node, and monocytes at the nucleotide level. The ligands for this receptor are CCL1 and CCL16.

Fig. 8: <span>Human thymus stained with anti-CCR8 antibody.</span>
Fig. 8: Human thymus stained with anti-CCR8 antibody. Cat.-No. AP15409PU-S.
 

CCR9

Fig. 9: <span>Immunofluorescence staining of mouse medulla of a mesenterial lymph node using CCR9 antibody AM01391SU-N (Blue: CD3, Green: IgA, Red: CCR9)</span>
Fig. 9: Immunofluorescence staining of mouse medulla of a mesenterial lymph node using CCR9 antibody AM01391SU-N (Blue: CD3, Green: IgA, Red: CCR9).
 

CCR10 is receptor for CCL27 and CCL28 that was originally called orphan receptor GPR2. CCR10 has been implicated in inflammation of the skin, and has been shown to recruit regulatory T cells to mucosal layers.

Fig. 10: <span>Immunohistochemical staining of tonsil using anti- CCR10 antibody SP4015P.</span>
Fig. 10: Immunohistochemical staining of tonsil using anti- CCR10 antibody SP4015P.
 

CCRL1 was originally designated CCR11 due to its ability to bind several CC chemokines (including CCL19, CCL21 and CCL25) and its structural similarity to chemokine receptors. However, due to the inability of this molecule to generate a signal following ligand interaction, it has been suggested that it is a scavenger receptor for chemokines and not a bonafide chemokine receptor. Thus CCRL1 should not be called CCR11 under the guidelines of the IUIS/WHO Subcommittee on Chemokine Nomenclature

Fig. 11: <span>Immunhistochemistry analysis in human heart (Formalin-fixed, Paraffin-embedded) using CCR11 / CCRL1 antibody Cat.-No. SP4097P.</span>
Fig. 11: Immunhistochemistry analysis in human heart (Formalin-fixed, Paraffin-embedded) using CCR11 / CCRL1 antibody Cat.-No. SP4097P.
 

Antibody Tools for Detection of Chemokine Receptors

Since Acris Antibodies was founded in 1998 we have increased the numbers of antibodies to chemokines and their receptors every year. Today we can offer an abundant antibody panel to chemokine receptors for various applications like Western blotting, immunohistochemistry, immunofluorescence, flow cytometry and other immunological detection methods.

Available antibody panel to C-C Chemokine Receptors

CCR1 antibody, CCR2 antibody, CCR3 antibody, CCR4 antibody, CCR5 antibody, CCR6 antibody, CCR7 antibody, CCR8 antibody, CCR9 antibody, CCR10 antibody, CCR11/CCRL1 antibody.

Literature

1. Rossi, D and Zlotnik, A (2000) The biology of chemokines and their receptors. Annu Rev Immunol. 18, 217-242, PubMed ID 20297045

2. Bagglioni, M. (1998) Chemokines and leukocyte traffic. Nature 392, 565-568 PubMed ID 98219079

For detailed background information on Chemokines and their receptors visit Horst Ibelgaufts\' COPE: Cytokines Online Pathfinder Encyclopaedia at www.copewithcytokines.de

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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  
SP4014P

CCR10 / GPR2 antibody

Figure 1. Immunohistochemical staining of thymus using anti-CCR10 antibody SP4014P.
Rabbit Aff - Purified Hu, Ms P
50 µg / €380.00
  Acris Antibodies GmbH
SP4015P

CCR10 / GPR2 (C-term) antibody

Immunohistochemical staining of tonsil using anti- CCR10 antibody SP4015P Rabbit Aff - Purified Hu P
50 µg / €380.00
  Acris Antibodies GmbH
SP7036P

CCR10 / GPR2 antibody

Western blot analysis for CCR10 using SP7036P at 2 µg/ml dilution against 10 ug of MCF-7 (a human breast cancer cell line) cell lysate.
Rabbit Purified Hu, Ms C, F, WB
0.1 mg / €330.00
  Acris Antibodies GmbH
SP4029P

CCR11 / CCRL1 antibody

Immunohistochemical staining of Heart, Cardiac myocytes using anti- CCRL1 antibody SP4029 Rabbit Aff - Purified Hu, Ms, Rt P
50 µg / €380.00
  Acris Antibodies GmbH
SP4097P

CCR11 / CCRL1 antibody

Immunhistochemistry analysis in human spleen (Formalin-fixed, Paraffin-embedded) using CCR11 / CCRL1 antibody Cat.-No. SP4097P. Rabbit Aff - Purified Hu P
50 µg / €380.00
  Acris Antibodies GmbH
AM26437AF-N

CD191 / CCR1 antibody

CD191 / CCR1 Mouse IgG1 141-2 Azide Free Hu F
0.1 mg / €320.00
  Acris Antibodies GmbH
AM26437RP-N

CD191 / CCR1 antibody

Flow cytometric analysis of CCR1 expression on human peripheral blood lymphocytes (left) and peripheral monocytes (right). Shaded histogram indicates the reaction of isotypic control to the cells. Open histogram indicates the reaction of AM26437RP-N to the cells. Mouse IgG1 141-2 PE Hu F
50 Tests / €330.00
  Acris Antibodies GmbH
AP08430PU-N

CD191 / CCR1 antibody

Lung: Formalin-Fixed Paraffin-Embedded (FFPE) 
  
Rabbit IgG Aff - Purified Hu, Ms P, WB
50 µg / €380.00
  Acris Antibodies GmbH
SP4001P

CD191 / CCR1 (Extracell. Dom.) antibody

Immunohistochemical staining of spleen lymphocytes using anti- CCR1 antibody SP4001P Rabbit Aff - Purified Hu, Ms P
50 µg / €380.00
  Acris Antibodies GmbH
SP7035P

CD191 / CCR1 antibody

Image 1: Surface staining of mouse primary epidermal cells (B-6 mice, 30wks) with CL 11-PE, CD11c PE-Cy5, and CCR1 (SP7035P) with CY5 goat anti rabbit secondary. Image 2: CD11c+ CL II+ cells (Langerans), stained with rabbit isotype control (shaded histogram) were analyzed for CCR1 at 5 µl (blue histogram) and 10 µl (green histogram). Data kindly provided by Corey Parlet, University of Iowa. Rabbit IgG Purified Hu, Ms F, WB
0.1 mg / €330.00
  Acris Antibodies GmbH
AP26344PU-N

CD192 / CCR2 antibody

AP26344PU-N: Western blot analysis of CCR2 expression in lysate from rat kidney (Lane 1), 3T3 cells (Lane 2) and Jurkat cells (Lane 3). Rabbit Aff - Purified Hu, Ms, Rt WB
0.1 mg / €340.00
  Acris Antibodies GmbH
SP4086P

CD192 / CCR2 antibody

Immunohistochemical staining of Spleen (Lymphocytes) using anti- CCR2 antibody SP4086P Rabbit Aff - Purified Hu P
50 µg / €380.00
  Acris Antibodies GmbH
AM26451AF-N

CD193 / CCR3 antibody

Flow cytometric analysis of CD193/CCR3 expression on transfectant. Open histogram indicates the reaction of isotypic control to the cells. Shaded histogram indicates the reaction of AM26451AF-N to the cells. Mouse IgG1 444-11 Azide Free Hu F
0.1 mg / €320.00
  Acris Antibodies GmbH
AM26451FC-N

CD193 / CCR3 antibody

Flow cytometric analysis of CD193 expression on transfectant. Open histogram indicates the reaction of isotypic control to the cells. Shaded histogram indicates the reaction of AM26451FC-N to the cells. Mouse IgG1 444-11 FITC Hu F
1 ml / €320.00
  Acris Antibodies GmbH
AM26451RP-N

CD193 / CCR3 antibody

Flow cytometricanalysis of CD193 /CCR3 expression on transfectant. Open histogram indicates the reaction of isotypic control to the cells. Shaded histogram indicates the reaction of AM26451RP-N to the cells. Mouse IgG1 444-11 PE Hu F
50 Tests / €320.00
  Acris Antibodies GmbH
AP30215CP-N

CD193 / CCR3 Control Peptide

CD193 / CCR3
50 µg / €130.00
  Acris Antibodies GmbH
AP30215PU-N

CD193 / CCR3 (C-term) antibody

Western blot analysis of CCR3 in human spleen tissue lysates with AP30215PU-N CCR3 antibody at 1 (lane 1) and 2 μg/ml (lane 2), and 2 μg/ml in the presence of blocking peptide (lane 3). Rabbit IgG Aff - Purified Hu, Ms, Rt E, P, WB
0.1 mg / €290.00
  Acris Antibodies GmbH
SP4002P

CD193 / CCR3 antibody

Immunohistochemical staining of  nasal mucosa (allergic rhinitis) using anti- CCR3 antibody SP4002P Rabbit Aff - Purified Hu, Ms P
50 µg / €380.00
  Acris Antibodies GmbH
SP4087P

CD193 / CCR3 antibody

Immunohistochemical staining of Nasal mucosa (Allergic Rhinitis, Eosinophils) using anti- CCR3 antibody SP4087P Rabbit Aff - Purified Hu P
50 µg / €380.00
  Acris Antibodies GmbH
SP7038P

CD193 / CCR3 antibody

Western blot analysis of 10 µg of total cell lysate from Daudi cells with SP7038P at 2 µg/ml dilution.
Rabbit Purified Hu, Ms, Rt WB
0.1 mg / €330.00
  Acris Antibodies GmbH

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