General description
Adhesion G protein-coupled receptor E5 (UniProt: P48960; also known as Leukocyte antigen CD97, CD97) is encoded by the ADGRE5 (also known as CD97) gene (Gene ID: 976) in human. CD97 is a multi-pass, homodimeric, membrane glycoprotein of the G-protein coupled receptor 2 family that is synthesized with a signal peptide (aa 1-20), which is subsequently cleaved off to produce the mature protein that contains four extracellular domains, seven transmembrane domains, and four cytoplasmic domains. The extracellular domain contains five EGF-like domains. The second and third EGF-like domains contain calcium binding sites, which are important in certain ligand interactions that are calcium-dependent. The fourth EGF-like domain mediates binding of chondroitin sulfate. The extracellular domain also contains one Arg-Gly-Asp (RGD) motif that serves as a recognition site for integrin binding. CD97 levels are quickly upregulated upon activation of most leukocytes. It is involved in cell adhesion and migration through interactions with cell surface proteins and components of the extracellular matrix. Three ligands for CD97 have been recognized: CD55 that serves as a negative regulator of complement cascade; chondroitin sulfate, and integrin 5β1. CD97 is broadly expressed and is found on most hematopoietic cells, including activated lymphocytes, monocytes, macrophages, dendritic cells, and granulocytes. Its expression is also observed on smooth muscle cells. Higher expression of CD97 is reported in thyroid, colorectal, esophageal, and pancreatic carcinomas. Its expression levels correlate with dedifferentiation, migration, and invasion in tumor cells. Three isoforms of CD97 have been described that are produced by alternative splicing. (Ref.: Safaee, M., et al. (2013). Int. J. Oncology. 43(5); 1343-1350; Steinert, M., et al. (2002). Am. J. Pathol. 161(5); 1657-1667; Kwakkenbos, MJ., et al. (2002). J. leukocyte Biol. 71(5); 854-862; Hamann, J., et al. (1996). J. Exp. Med. 184(3); 1185-1189).
Immunogen
NIH3T3 cells stably expressing human CD97.
Application
Quality Control Testing
Evaluated by Flow Cytometry in THP-1 cells.
Flow Cytometry Analysis: 1.0 µg of this antibody detected CD97 in one million THP-1 cells.
Tested Applications
Flow Cytometry Analysis: A representative lot detected CD97 in Flow Cytometry applications (Hamann, J., et al. (1996). 184(3):1185-9; Kwakkenbos, M.J., et al. (2002). J Leukoc Biol. 71(5):854-62).
Inhibition Assay: A representative lot of this antibody inhibited erythrocyte adhesion to CD97-transfected COS cells.(Hamann, J., et al. (1996). 184(3):1185-9; Hamann, J., et al. (1998). Eur J Immunol.;28(5):1701-7).
Immunoprecipitation Analysis: A representative lot immunoprecipitated CD97 in Immunoprecipitation applications (Hamann, J., et al. (1996). 184(3):1185-9).
Immunohistochemistry Applications: A representative lot detected CD97 in Immunohistochemistry applications (Hamann, J., et al. (1999). Arthritis Rheum. 42(4):650-8; Steinert, M., et al. (2002). Am J Pathol. 161(5):1657-67).
Immunofluorescence Analysis: A representative lot detected CD97 in Immunofluorescence applications (Hamann, J., et al. (1998). Eur J Immunol. 28(5):1701-7; Hamann, J., et al. (1999). Arthritis Rheum. 42(4):650-8; Steinert, M., et al. (2002). Am J Pathol. 161(5):1657-67).
ELISA Analysis: A representative lot detected CD97 in ELISA applications (Hamann, J., et al. (1999). Arthritis Rheum. 42(4):650-8; Steinert, M., et al. (2002). Am J Pathol. 161(5):1657-67).
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Biochem/physiol Actions
Clone CLB‐CD97/1 is a mouse monoclonal antibody that detects CD97 (Adhesion G protein-coupled receptor E5).
Physical form
Purified mouse monoclonal antibody IgG2a in PBS without preservatives.
Preparation Note
1.0 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
