Human CD97 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human CD97 in ELISAs. In sandwich immunoassays, no cross-reactivity or interference with recombinant human (rh) CD55, rhEGF, rhEMR2, or recombinant mouse CD97 is observed.
  • Source
    Monoclonal Mouse IgG2B Clone # 380905
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human CD97
    Gln21-Gln398
    Accession # NP_001775.2
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
    • Human CD97 Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    2-8 µg/mL 
    Human CD97 Antibody (Catalog # MAB25291)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Human CD97 Biotinylated Antibody (Catalog # BAF2529)
  • ELISA Standard
     
    Recombinant Human CD97 Protein, CF (Catalog # 2529-CD)
  • Neutralization
    Measured by its ability to neutralize CD97-mediated adhesion of human red blood cells. Hamann, J. et al. (1996) J. Exp. Med. 184:1185. The Neutralization Dose (ND50) is typically 0.25-1.25 µg/mL in the presence of 4 µg/mL Recombinant Human CD97.
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.
Data Examples
Cell Adhesion Mediated by CD97 and Neutralization by Human CD97 Antibody. Recombinant Human CD97 (Catalog # 2529-CD), immobil­ized onto a microplate, supports the adhesion of human red blood cells in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Human CD97 (4 µg/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human CD97 Monoclonal Antibody (Catalog # MAB25291). The ND50 is typically 0.25-1.25 µg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & 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.
Background: CD97

CD97 is a 95‑100 kDa member of a protein group known as the LNB-TM7 protein family that evolved from genes of the secretin receptor superfamily (1‑3). Molecules in this family are unique hybrid structures consisting of EGF-like modules coupled to class B G-protein 7-transmembrane (TM) domains by a glycosylated (mucin) stalk. Human CD97 is synthesized as an 835 amino acid (aa) precursor that contains a 20 aa signal sequence, a 532 aa extracellular domain (ECD), a 238 aa “transmembrane” region that includes seven TM segments, and a 45 aa cytoplasmic tail (4). Within the 532 aa ECD, the first 236 aa contains five EGF-like domains, the C-terminal four of which bind calcium, and a juxtamembrane 296 aa RGD-containing mucin stalk (4, 5). The stalk is both glycosylated and proteolytically cleaved (at aa 530) to create a noncovalently linked 65‑70 kDa glycosylated extracellular alpha -subunit and a 28 kDa 7-TM membrane-bound beta -subunit (4). There are two known alternate splice forms in human. Isoform # 1 contains four EGF-like domains (domain # 1, 2, 3 and 5), while isoform # 2 contains three EGF-like domains (domain # 1, 2 and 5) (1, 4, 6). The ECD in isoform 1 is 60 kDa in size, while the ECD in isoform 2 is 55 kDa in size (native molecular weight). The five EGF-like domain region in human shares 55% aa sequence identity with that in mouse. Cells known to express CD97 include monocytes, macrophages, T cells, select B cells, dendritic cells and, potentially, vascular and visceral smooth muscle cells (1, 7). There are at least two ligands for CD97. One is chrondroitin sulfate that binds only to the full-length (five domain) form of CD97. Binding is dependent on the presence of EGF-like domain # 4 (3). The second ligand for CD97 is CD55, a GPI-linked cell surface molecule with short consensus repeats that regulates complement activation on cell surfaces (1, 5, 7). CD97 EGF-like domains # 1 and 2 bind CD55 while domain # 5 stabilizes the CD97 molecule. The shortest CD97 isoform shows the strongest binding to CD55 (7, 8).

  • References:
    1. McKnight, A.J. and S. Gordon (1998) J. Leukoc. Biol. 63:271.
    2. Stacey, M. et al. (2000) Trends Biochem. Sci. 25:284. 
    3. Stacey, M. et al. (2003) Blood 102:2916. 
    4. Gray, J.X. et al. (1996) J. Immunol. 157:5438. 
    5. Lin, H-H. et al. (2001) J. Biol. Chem. 276:24160.
    6. Hamann, J. et al. (1995) J. Immunol. 155:1942.
    7. Jaspars, L.H. et al. (2001) Tissue Antigens 57:325.
    8. Hamann, J. et al. (1996) J. Exp. Med. 184:1185.
  • Entrez Gene IDs:
    976 (Human); 26364 (Mouse)
  • Alternate Names:
    CD97
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