About InVivoMAb anti-mouse CCR5 (CD195) The C5Mab-2 monoclonal antibody reacts with mouse CC chemokine receptor type-5 (CCR5), also known as MCP-1 receptor, CKR5, and CD192. CCR5 is a seven-pass transmembrane protein from the GPCR family, and is expressed on dendritic cells (DCs), T cells (including Tregs), macrophages, eosinophils, NK cells, myeloid-derived suppressor cells (MDSCs), and some non-hematopoietic cells such as epithelial cells, endothelial cells, fibroblasts (including vascular smooth muscle), and microglia. The natural agonistic, or activating, ligands of CCR5 include CCL3/MIP-1 alpha, CCL3L1, CCL4 (MIP-1 beta), CCL5/RANTES, CCL8, CCL11, CCL13, and CCL16. Notably, CCL7/MCP-3 binds to CCR5 as a natural antagonist/deactivating ligand, thereby blocking receptor signaling and influencing inflammation and immune responses in a complex and context-dependent manner. CCR5 receptor activation regulates the migration and activation of lymphocytes, immune surveillance, tumorigenesis, and inflammation, including the pathogenesis of inflammatory diseases. To facilitate the recruitment of the immune cells to inflammation sites, CCR5 directs the process of chemotaxis (immune cell migration) along the chemokine gradient. In neurobiology, CCR5 acts as a suppressor of learning, memory, neuronal plasticity, and synaptic connections in the brain. CCR5 is well documented to act as a common coreceptor for HIV-1, and CCR5 antagonists are reported to provide protection against HIV-1. In vivo studies with mouse models of traumatic brain injury (TBI) have linked CCR5 knockdown to reduced learning deficit and improvement of cognitive function. In view of the pleiotropic effects of CCR5, its antagonists, including monoclonal antibodies, have emerged as promising tools for experimental therapeutics of tumors and inflammatory diseases. InVivoMAb anti-mouse CCR5 (CD195) Specifications IsotypeRat IgG2b, κ Recommended Isotype Control(s)InVivoMAb rat IgG2b isotype control, anti-keyhole limpet hemocyanin Recommended Dilution BufferInVivoPure pH 7.0 Dilution Buffer ImmunogenMouse CCR5 expressing CHO-K1 cells Reported ApplicationsFlow cytometry For details on in vivo applications, please contact technicalservice@bioxcell.com FormulationPBS, pH 7.0 Contains no stabilizers or preservatives Endotoxin<2EU/mg (<0.002EU/μg) Determined by LAL gel clotting assay Purity>95% Determined by SDS-PAGE Sterility0.2 μm filtered ProductionPurified from cell culture supernatant in an animal-free facility PurificationProtein G Molecular Weight150 kDa StorageThe antibody solution should be stored at the stock concentration at 4°C. Do not freeze. Application ReferencesInVivoMAb anti-mouse CCR5 (CD195) (CLONE: C5Mab-2)Suzuki H, Tanaka T, Li G, Ouchida T, Kaneko MK, Kato Y (2024). "Development of a Sensitive Anti-Mouse CCR5 Monoclonal Antibody for Flow Cytometry" Monoclon Antib Immunodiagn Immunother 43(4):96-100. PubMedC-C chemokine receptor 5 (CCR5), a member of the G protein-coupled receptor family, is the most common coreceptor for the human immunodeficiency virus type 1. CCR5 is also involved in the pathogenesis of tumors and inflammatory diseases. The CCR5 antagonists including monoclonal antibodies (mAbs) have been developed and evaluated in clinical trials. In this study, we developed novel mAbs for mouse CCR5 (mCCR5) using the Cell-Based Immunization and Screening (CBIS) method. One of the established anti-mCCR5 mAbs, C5Mab-2 (rat IgG2b, kappa), reacted with mCCR5-overexpressed Chinese hamster ovary-K1 (CHO/mCCR5) and an endogenously mCCR5-expressing cell line (L1210) by flow cytometry. Using flow cytometry, the dissociation constant (KD) of C5Mab-2 for CHO/mCCR5 was determined as 4.3 × 10-8 M. These results indicated that C5Mab-2 is useful for the detection of mCCR5 in flow cytometry and may be applicable to obtain the proof of concept in preclinical studies.Ubukata R, Suzuki H, Tanaka T, Li G, Kaneko MK, Kato Y (2024). "Development of Sensitive Anti-Mouse CCR5 Monoclonal Antibodies Using the N-Terminal Peptide Immunization" Monoclon Antib Immunodiagn Immunother 43(4):112-118. PubMedOne of the G protein-coupled receptors, C-C chemokine receptor 5 (CCR5), is an important regulator for the activation of T and B lymphocytes, dendritic cells, natural killer cells, and macrophages. Upon binding to its ligands, CCR5 activates downstream signaling, which is an important regulator in the innate and adaptive immune response through the promotion of lymphocyte migration and the secretion of proinflammatory cytokines. Anti-CCR5 monoclonal antibodies (mAbs) have been developed and evaluated in clinical trials for tumors and inflammatory diseases. In this study, we developed novel mAbs for mouse CCR5 (mCCR5) using the N-terminal peptide immunization. Among the established anti-mCCR5 mAbs, C5Mab-4 (rat IgG2a, kappa) and C5Mab-8 (rat IgG1, kappa), recognized mCCR5-overexpressing Chinese hamster ovary-K1 (CHO/mCCR5) and an endogenously mCCR5-expressing cell line (L1210) by flow cytometry. The dissociation constant (KD) values of C5Mab-4 and C5Mab-8 for CHO/mCCR5 were determined as 3.5 × 10-8 M and 7.3 × 10-9 M, respectively. Furthermore, both C5Mab-4 and C5Mab-8 could detect mCCR5 by western blotting. These results indicated that C5Mab-4 and C5Mab-8 are useful for detecting mCCR5 by flow cytometry and western blotting and provide a possibility to obtain the proof of concept in preclinical studies.
