Search results for: Rat monoclonal anti mouse CXCR6 Anti-Mouse antibodies
#36301196 2022/10/25 To Up
Epitope Mapping of an Anti-Mouse CXCR6 Monoclonal Antibody (CxMab-1) Using the 2 × Alanine Scanning Method.
The CXC chemokine receptor 6 (CXCR6) is a member of the G protein-coupled receptor family that is highly expressed in helper T type 1 cells, cytotoxic T lymphocytes (CTLs), and natural killer cells. CXCR6 plays critical roles in local expansion of effector-like CTLs in tumor microenvironment to potentiate the antitumor response. Therefore, the development of anti-CXCR6 monoclonal antibodies (mAbs) is essential to evaluate the immune microenvironment of tumors. Using N-terminal peptide immunization, we previously developed an anti-mouse CXCR6 (mCXCR6) mAb, CxMab-1 (rat IgG1, kappa) , which is useful for flow cytometry and western blotting. In this study, we determined the critical epitope of CxMab-1 by enzyme-linked immunosorbent assay (ELISA) using the 1 × alanine scanning (1 × Ala-scan) method or the 2 × alanine scanning (2 × Ala-scan) method. Although we first performed ELISA by 1 × Ala-scan using one alanine-substituted peptides of mCXCR6 N-terminal domain (amino acids 1-20), we could not identify the CxMab-1 epitope. We next performed ELISA by 2 × Ala-scan using two alanine (or glycine) residues-substituted peptides of mCXCR6 N-terminal domain, and found that CxMab-1 did not recognize S8A-A9G, A9G-L10A, L10A-Y11A, and G13A-H14A of the mCXCR6 N-terminal peptide. The results indicate that the binding epitope of CxMab-1 includes Ser8, Ala9, Leu10, Tyr11, Gly13, and His14 of mCXCR6. Therefore, we could demonstrate that the 2 × Ala scan method is useful for determining the critical epitope of mAbs.Yu Isoda, Tomohiro Tanaka, Hiroyuki Suzuki, Teizo Asano, Takuro Nakamura, Miyuki Yanaka, Saori Handa, Yu Komatsu, Saori Okuno, Nozomi Takahashi, Yuki Okada, Hiyori Kobayashi, Guanjie Li, Ren Nanamiya, Nohara Goto, Nami Tateyama, Takeo Yoshikawa, Mika K Kaneko, Yukinari Kato
1104 related Products with: Epitope Mapping of an Anti-Mouse CXCR6 Monoclonal Antibody (CxMab-1) Using the 2 × Alanine Scanning Method.
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#35736626 // To Up
CxMab-1: A Novel Anti-Mouse CXCR6 Monoclonal Antibody Established by N-Terminal Peptide Immunization.
The CXC chemokine receptor 6 (CXCR6) is a member of the G protein-coupled receptor family that is highly expressed in helper T type 1 cells, natural killer cells, cytotoxic T lymphocytes, and various type of cells in tumor microenvironment (TME). CXCR6 has been proposed as a therapeutic target against tumors through regulation of the tumor TME. In this study, we developed specific and sensitive monoclonal antibodies (mAbs) for mouse CXCR6 (mCXCR6), which are useful for flow cytometry and Western blotting by N-terminal peptide immunization into rat. The established anti-mCXCR6 mAb, CxMab-1 (rat IgG, kappa), reacted with not only mCXCR6-overexpressed Chinese hamster ovary-K1 (CHO/mCXCR6) but also mCXCR6-endogenously expressed cell lines, such as P388 (mouse lymphoid neoplasm) and J774-1 (mouse macrophage-like) through flow cytometry. Kinetic analyses using flow cytometry indicated that the dissociation constants () of CxMab-1 for CHO/mCXCR6, P388, and J774-1 cells were 1.7 × 10 M, 3.4 × 10 M, and 3.8 × 10 M, respectively. Furthermore, CxMab-1 could detect endogenous mCXCR6 in P388 and J774-1 cells by Western blotting. These results indicated that CxMab-1 is useful for detecting mCXCR6 by flow cytometry and Western blotting, and provides a possibility for targeting CXCR6-expressing cells experiments.Kaishi Kitamura, Hiroyuki Suzuki, Mika K Kaneko, Yukinari Kato
2185 related Products with: CxMab-1: A Novel Anti-Mouse CXCR6 Monoclonal Antibody Established by N-Terminal Peptide Immunization.
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