Rabbit anti-Human CD97 mAb for FC - P48960

Core Product Specifications and Parameters

Biological Background: CD97 Function and Localization

• CD97, also known as Adhesion G protein-coupled receptor E5 or Leukocyte antigen CD97, is a member of the adhesion GPCR family characterized by an extended extracellular N-terminus containing multiple EGF-like repeats.
• It functions as a receptor involved in both cell adhesion and signaling, playing an essential role in leukocyte migration.
• The protein is expressed as transmembrane and soluble isoforms; alternative splicing generates variants with different numbers of EGF-like domains.
• Subcellular localization: primarily found on the cell membrane, but also secreted into the extracellular space.
• Broadly expressed on most hematopoietic cells (activated lymphocytes, monocytes, macrophages, dendritic cells, granulocytes) and abundant in smooth muscle cells; also found in certain carcinomas (thyroid, colorectal, gastric, esophageal, pancreatic).
• Expression is upregulated under inflammatory conditions, such as in multiple sclerosis CNS and rheumatoid arthritis synovial tissue, with increased soluble CD97 detectable in synovial fluid.
• Post-translational modifications include glycosylation, disulfide bonds, and phosphorylation; the extracellular domain contains EGF-like calcium-binding sites.

Experimental Guidance and Technical Tips

• The immunogen used was a recombinant protein corresponding to amino acids 21-398 of human CD97, encompassing the extracellular domain. This design likely yields antibodies recognizing surface-accessible epitopes.
• For flow cytometry, use standard staining protocols with appropriate isotype controls; titrate the antibody to determine optimal working concentration.
• Validate reactivity in relevant human hematopoietic or carcinoma cell lines known to express CD97 (e.g., leukocytes, smooth muscle cell lines).
• Note that CD97 can be shed, so soluble receptor may compete in certain assays; consider using fresh cells or blocking with recombinant protein if necessary.
• Avoid relying on molecular weight determination by western blot as this antibody is recommended for FC only; verify specificity by staining cells with known CD97 expression patterns.

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