Detection of CD99 in Human PBMCs by Flow Cytometry.
Human peripheral blood mononuclear cells were stained with Goat Anti-Human CD99 Fluorescein‑conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # FAB3968F, filled histogram) or isotype control antibody (Catalog # IC108F, open histogram). View our protocol for Staining Membrane-associated Proteins.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
CD99 (also named MIC2, E2 and thymic leukemia antigen) is the founding member of the CD99 family of molecules. The CD99 family contains four members; CD99, CD99L2, XG and the pseudogene CD99L1 (1, 2, 3). Native human CD99 is 32 kDa in size and exists as a type I transmembrane glycoprotein. This is referred to as the long, or type I isoform. It is synthesized as a 185 amino acid (aa) precursor that contains a 22 aa signal sequence, a 100 aa extracellular domain (ECD), a 25 aa transmembrane segment, and a 38 aa cytoplasmic region (4). The ECD contains no identifiable motifs, N‑linked glycosylation sites, or cysteine residues; it does possess sites for O-linked glycosylation. The cytoplasmic region, albeit short, does have signal transduction capability (5). There are apparently multiple isoforms for human CD99. One shows a 16 aa deletion in the ECD (aa 34‑49), a second shows a 38 aa deletion in the cytoplasmic region (aa 122‑159), and a third exhibits a three aa truncation at the C-terminus (6, 7, 8). The best studied isoform shows an Asp‑Gly substitution for the C‑terminal 27 amino acids. This is referred to as the 28 kDa type II isoform (9). The type I and II isoforms have distinctive signal transduction pathways (FAK-src for type I; PI3K plus src-ERK1/2 for type II), and mediate clearly different biological outcomes (5, 9, 10). The two numbered isoforms may or may not co‑exist on the same cells. Peripheral T cells have only the long isoform, while double-positive thymocytes express both isotypes. What is unclear is the monomeric vs. dimeric status of CD99. In mouse, CD99 reportedly forms disulfide-linked homodimers (11). In human, however, CD99 is reportedly monomeric if only a type I isoform, and a covalent heterodimer if co‑expressing type I and II isoforms (12, 13). Cells known to express CD99 include fibroblasts, neutrophils, T cells, double-positive thymocytes, CD34+ stem cells, monocytes and endothelial cells (2, 12, 14, 15). Homophilic interaction between CD99 on the neutrophil and CD99 on the endothelial cell regulates the transendothelial migration of neutrophils during inflammation (16). Over aa 23-122, human CD99 shares only 43% aa sequence identity with mouse CD99 (17).
Wilson, M.D. et al. (2006) Physiol. Genomics 27:201.
Petri, B. and M.G. Bixel (2006) FEBS J. 273:4399.
Suh, Y.H. et al. (2003) Gene 307:63.
Gelin, C. et al. (1989) EMBO J. 8:3253.
Byun, H-J. et al. (2006) J. Biol. Chem. 281:34833.
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