Human/Mouse CTLA-4 Antibody Summary
Accession # P16410
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human and Mouse CTLA‑4 by Western Blot. Western blot shows lysates of NS0 mouse myeloma cell line transfected with with Human CTLA‑4 and mouse lymph node tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse CTLA‑4 Monoclonal Antibody (Catalog # MAB386) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for CTLA‑4 at approximately 52 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Inhibition of Anti-CD3 Induced IFN-gamma Secretion by Human/Mouse CTLA‑4 Antibody. Anti-Human/Mouse CTLA-4 Monoclonal Antibody (Catalog # MAB386) inhibits IFN-gamma secretion in human peripheral blood mononuclear cells (PBMC) induced by immobilized Mouse Anti-Human CD3 epsilon Monoclonal Antibody (Catalog # MAB100)coatedat 1 μg/mL (100 μL/well) in a dose-dependent manner (green line) as measured by the Human IFN-gamma Quantikine ELISA Kit (Catalog # DIF50). The ED50 is typically 0.05-0.4 μg/mL.
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
CTLA-4 (Cytotoxic T-lymphocyte Associated protein‑4, designated CD152), is a type I transmembrane T cell inhibitory molecule that is a member of the Ig superfamily (1, 2). Human or mouse CTLA-4 cDNA encodes 223 amino acids (aa) including a 35 aa signal sequence, a 126 aa extracellular domain (ECD) with one Ig-like V-type domain, a 21 aa transmembrane (TM) sequence, and a 41 aa cytoplasmic sequence. It is found as a covalent homodimer of 41-43 kDa (2) Within the ECD, human CTLA-4 shares 68%, 71% and 83‑86% aa sequence identity with mouse, rat and porcine/bovine/rabbit/feline/canine CTLA-4, respectively. A 174 aa form that lacks TM and cytoplasmic sequences (sCTLA-4) is possibly secreted (3-5). Isoforms of 56-79 aa that mainly contain parts of the cytoplasmic domain are reported. In mouse, an isoform lacking the Ig-like domain has ligand-independent inhibitory activity and is termed liCTLA-4 (6). CD28, which is structurally related to CTLA-4, is constitutively expressed on naïve T cells and promotes T cell activation when engaged by B7-2 on antigen-presenting cells (APC) within the immunological synapse (IS) (1, 7, 8). In contrast, CTLA-4 is recruited from intracellular vesicles to the IS beginning 1-2 days after T cell activation (2, 7, 8). It forms a linear lattice with B7-1 on APC, inducing negative regulatory signals and ending T cell activation (9). Abatacept, a therapeutic human CTLA-4-Ig fusion protein (trade name Orencia), competes with CD28 for B7-1 and B7-2 binding and has been used to antagonize T cell activation in autoimmune conditions and to enhance transplant survival (10). Mice deleted for CTLA-4 show no abnormalities until after birth, but then develop lethal autoimmune reactions due to continued T cell activation and poor control by regulatory T cells, which constitutively express CTLA-4 in wild-type mice and humans (11-13).
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