|Detection of Human and Mouse CD27/TNFRSF7 by Western Blot. Western blot shows lysates of human lymph node tissue and EL‑4 mouse lymphoblast cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human/Mouse CD27/TNFRSF7 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF574) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CD27/TNFRSF7 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|CD27/TNFRSF7 in Mouse Splenocytes. CD27/TNFRSF7 was detected in immersion fixed mouse splenocytes using Goat Anti-Human/Mouse CD27/TNFRSF7 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF574) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.|
CD27 is a lymphocyte-specific member of the tumor necrosis factor receptor superfamily (TNFRSF) and is designated TNFRSF7 (1, 2). Mouse CD27 cDNA encodes a 250 amino acid (aa) residue type I transmembrane protein with a 20 aa putative signal peptide, a 162 aa extracellular region containing three TNFR cysteine-rich repeats, a 21 aa transmembrane domain and a 47 aa cytoplasmic region (3). Mouse and human CD27 share approximately 65% amino acid identity. CD27 exists as homodimers on the cell surface via an extracellular disulfide bond in the membrane-proximal region. A soluble form of CD27 is also produced during the immune response and is found in various body fluids (4). CD27 is expressed on subsets of T and B cells. The expression of CD27 is upregulated upon T-cell activation. Although CD27 appears to be a marker for human memory B cells, it is only expressed in a small population of mouse B cells in germinal centers and at sites of B cell stimulation, suggesting that mouse CD27 may be a marker for activated B cells (5). CD27 interacts with CD27 ligand (also named CD70 and TNFSF7), which is a member of the TNF ligand superfamily. Ligation of CD27 on T cells provides costimulatory signals that are required for T cell proliferation, clonal expansion and the promotion of effector T cell formation (1, 2). Ligation of CD27 on B cells has been shown to inhibit terminal differentiation of activated mouse B cells into plasma cells and enhances commitment to memory B cell responses (5).