CD27/TNFRSF7 Antibody [Unconjugated]

Western blot shows lysates of Raji human Burkitt's lymphoma cell line, Daudi human Burkitt's lymphoma cell line, Ramos human Burkitt's lymphoma cell line, and human tonsil tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human CD27/TNFRSF7 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF382) 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 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human CD27/TNFRSF7 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF382) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes in germinal center. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Simple Western lane view shows lysates of human tonsil tissue, loaded at 0.2 mg/mL. A specific band was detected for CD27/TNFRSF7 at approximately 49 kDa (as indicated) using 20 µg/mL of Goat Anti-Human CD27/TNFRSF7 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF382) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

In the presence of sub-optimal amounts of Mouse CD3 epsilon Monoclonal Anti­body (Catalog # MAB484) and Recombinant Mouse CD27 Ligand (10 µg/mL, Catalog # 783-CL), Recom­binant Human CD27 Fc Chimera (Catalog # 382-CD) inhibits proliferation in mouse splenic T cells in a dose-dependent manner (orange line). Under these conditions, inhibition of proliferation elicited by Recombinant Human CD27 Fc Chimera (3 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human CD27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF382). The ND50 is typically 2.25-9.0 µg/mL.

• Reactivity: Hu
• Applications: WB, Simple Western, Flow, IHC, B/N, CyTOF-ready
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

CD27/TNFRSF7 Antibody [Unconjugated] Summary

• Immunogen: S. frugiperda insect ovarian cell line Sf 21-derived recombinant human CD27
  Thr21-Ile192
  Accession # P26842
• Specificity: Detects human CD27 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CD27
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunohistochemistry 3-15 ug/mL
  • Neutralization 2.25-9.0 ug/mL
  • Simple Western 20 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CD27/TNFRSF7 Antibody [Unconjugated]

• CD27 antigen
• CD27 molecule
• CD27
• MGC20393
• S152CD27L receptor
• T cell activation antigen CD27
• T cell activation antigen S152
• T14
• TNFRSF7
• TNFRSF7T-cell activation antigen CD27
• Tp55
• Tumor necrosis factor receptor superfamily member 7
• tumor necrosis factor receptor superfamily, member 7

Background

Human CD27 is a lymphocyte-specific member of the TNF receptor superfamily. CD27 is expressed on a subset of human thymocytes and on the majority of mature T cells. CD27 expression is up-regulated after TCR stimulation. Within the CD4⁺ compartment, it is preferentially expressed on CD45RA⁺ cells. In contrast, it is preferentially expressed on CD45RO⁺ cells in the CD8⁺ compartment. CD27 also appeaars to be a potential marker for memory B cells. It exists as both a disulfide-linked dimer on the cell surface and as a soluble protein found in serum. Human CD27 is a 260 amino acid (aa) protein with a 20 aa signal, a 173 aa extracellular domain, a 20 aa transmembrane domain, and a 47 aa cytoplasmic domain. The ligand for CD27 is CD70. CD70 is expressed on thymic stromal cells and a small subset of activated T cells. Additionally a subset of activated B cells express CD70. The CD27/CD70 interaction appears to be a weak costimulatory pathway involved in T cell and B cell immune response. CD27/CD70 interactions may be more involved in controlling the expansion phase of an immune response. This would be in contrast to B7/CD28 interactions, which are important for the activation phase of immune responses.

1. Camerini, D. et al. (1991) J. Immunol. 147:3165.
2. Loenen, W.A. et al. (1992) J. Immunol. 149:3937.
3. Lens, S.M.A. et al. (1998) Sem. Immunol. 10:491.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CD27/TNFRSF7 Antibody (AF382)(2)

Publications using AF382

• Vranjkovic A, Crawley AM, Gee K, Kumar A, Angel JB IL-7 decreases IL-7 receptor alpha (CD127) expression and induces the shedding of CD127 by human CD8+ T cells. Int. Immunol., 2007;19(12):1329-39. 2007-01-01 [PMID: 17956896] (Western Blot, Human)
• Yun LW, Decarlo AA, Hunter N Blockade of protease-activated receptors on T cells correlates with altered proteolysis of CD27 by gingipains of Porphyromonas gingivalis. Clin. Exp. Immunol., 2007;150(2):217-29. 2007-01-01 [PMID: 17937677] (ELISA Development, Human)

Bioinformatics

• Gene Symbol: CD27
• Uniprot:
  • P26842()