About InVivoMAb anti-mouse CCR2 (CD192) The C2Mab-6 monoclonal antibody reacts with mouse CC chemokine receptor type-2 (CCR2) also known as MCP-1 receptor and CD192. CCR2 is the primary receptor of C-C motif ligand 2 (CCL2) also known as monocyte chemoattractant protein-1 (MCP-1). CCR2 can also bind CCL7 and CCL12. CCR2 is a multi-pass transmembrane G protein-coupled receptor (GPCR) that is expressed on natural killer (NK) cells, macrophages, peripheral blood monocytes, and activated T cells, B cells, and immature dendritic cells (DCs). CCL2-CCR2 signaling is involved in the regulation of immune cells’ chemotaxis/migration, and during microbial infections, CCR2-positive macrophages and neutrophils drive innate immune responses. In neurobiology, CCR2 plays an important role in mediating peripheral nerve injury-induced neuropathic pain and facilitates the recruitment of macrophages/monocytes to the injury site following cerebral injury. The crosstalk between CCR2 on tumor-infiltrating immune and stromal cells and CCL2 on tumor cells is known to influence tumor growth, angiogenesis, and metastasis. CCR2 antagonists are often studied in relation to unwanted immune responses in inflammatory and autoimmune diseases, immunomodulation of the tumor microenvironment (TME) for enhancing anti-tumor immunity, and Treg function. The C2Mab-6 antibody is raised against and binds to the N-terminal sequence of CCR2, a region important for ligand recognition and binding. Cell-based screening has shown that the C2Mab-6 antibody has a strong affinity for both endogenous and exogenous mouse CCR2, indicating that this antibody can potentially be used to block CCR2 in in vivo preclinical studies. InVivoMAb anti-mouse CCR2 (CD192) Specifications IsotypeRat IgG1, κ Recommended Isotype Control(s)InVivoMAb rat IgG1 isotype control, anti-horseradish peroxidase Recommended Dilution BufferInVivoPure pH 7.0 Dilution Buffer ImmunogenThree KLH-conjugated synthetic peptides corresponding to the N-terminal sequence of mouse CCR2 Reported ApplicationsFlow cytometry ELISA 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 CCR2 (CD192) (CLONE: C2Mab-6)Tanaka T, Li G, Asano T, Saito M, Kaneko MK, Suzuki H, Kato Y (2022). "Development of a Novel Anti-Mouse CCR2 Monoclonal Antibody (C2Mab-6) by N-Terminal Peptide Immunization" Monoclon Antib Immunodiagn Immunother 41(2):80-86. PubMedThe CC chemokine receptor type-2 (CCR2) belongs to the G-protein-coupled receptor superfamily, expressed on the cell surface of immune cells and tumors. CCR2 binds to the CC motif chemokine 2/monocyte chemoattractant protein-1, a CC chemokine, which is produced by various cells, including immune-related cells and tumors. Therefore, the development of sensitive monoclonal antibodies (mAbs) for CCR2 has been desired for treatment and diagnosis. This study established a novel, specific, and sensitive anti-mouse CCR2 (mCCR2) mAb; C2Mab-6 (rat IgG1, kappa), using the mCCR2 synthetic peptide immunization method. C2Mab-6 reacted with mCCR2-overexpressed Chinese hamster ovary-K1 cells and L1210 (murine leukemia) cells, which express endogenous mCCR2 in flow cytometry. Furthermore, C2Mab-6 showed a high binding affinity for both cells. Hence, C2Mab-6 can be a useful tool for analyzing mCCR2-related biological responses, using flow cytometry.Tanaka T, Suzuki H, Asano T, Li G, Nanamiya R, Tateyama N, Isoda Y, Okada Y, Kobayashi H, Yoshikawa T, Kaneko MK, Kato Y (2022). "Epitope Mapping of an Anti-Mouse CCR2 Monoclonal Antibody (C2Mab-6) Using Enzyme-Linked Immunosorbent Assay" Monoclon Antib Immunodiagn Immunother 41(6):339-342. PubMedCC chemokine receptor type-2 (CCR2) is a member of the G protein-coupled receptors, and is mainly expressed on cell surface of immune cells. CCR2 binds to its ligand, C-C motif chemokine 2 (also named as monocyte chemoattractant protein-1), which involves in the tumor progression by modulating the tumor microenvironment. Therefore, the monoclonal antibody (mAb) targeting CCR2 could be one of the strategies for cancer treatment. In this study, we investigated the critical epitope of C2Mab-6, an anti-mouse CCR2 (mCCR2) mAb developed by N-terminal peptides immunization. We first performed enzyme-linked immunosorbent assay (ELISA) using N-terminal peptides of mCCR2 and demonstrated that C2Mab-6 recognizes 1-19 amino acids of mCCR2. We further performed ELISA using 20 alanine-substituted peptides of mCCR2. C2Mab-6 lost the reaction to the alanine-substituted peptides of D3A, N4A, M6A, P8A, Q9A, and F10A. These results indicate that the binding epitope of C2Mab-6 includes Asp3, Asn4, Met6, Pro8, Gln9, and Phe10 of mCCR2.
