Detection of CD229/SLAMF3 in Human PBMCs by Flow Cytometry.
Human peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human CD3 epsilon PE‑conjugated Monoclonal Antibody (Catalog # FAB100P) and either (A) Mouse Anti-Human CD229/SLAMF3 APC‑conjugated Monoclonal Antibody (Catalog # FAB1898A) or (B) Mouse IgG2A Allophycocyanin Isotype Control (Catalog # IC003A). 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.
CD229, also known as Ly9 and SLAMF3, is a 120 kDa type I transmembrane glycoprotein in the SLAM subgroup of the CD2 family (1). Mature human CD229 consists of a 407 amino acid (aa) extracellular domain (ECD) with two Ig‑like V-set and two Ig‑like truncated C2-set domains. It also shows a 22 aa transmembrane segment, and a 179 aa cytoplasmic domain with two immunoreceptor tyrosine-based switch motifs ITSMs (2, 3). The ECD of human CD229 shares 57%‑59% aa sequence identity with mouse and rat CD229. Within the first two Ig‑like domains that are common to all SLAM proteins, human CD229 shares 24%‑39% aa sequence identity with human 2B4, BLAME, CD2F-10, CD84, CRACC, NTB-A, and SLAM. Alternate splicing generates two additional isoforms that lack the juxtamembrane Ig‑like domain or short cytoplasmic region (2). CD229 is expressed on T and B cells, thymocytes, and more weakly on NK cells (2‑6). Homophilic binding between CD229 molecules is mediated by the N-terminal Ig‑like domain (7). Human and mouse CD229 exhibit cross-species binding (7). Antigen stimulation of lymphocytes induces CD229 clustering to sites of T cell‑B cell contact (7). Two tyrosines in the cytoplasmic domain are required for association of CD229 with the adaptor proteins AP-2 (μ2 chain) and GRB-2 and both are required for CD229 internalization (8, 9). In addition, there are two ITSMs which mediate phosphorylation-dependent CD229 association with SAP and SHP-2 (10). These four tyrosine-containing motifs are intact in the described CD229 splice variants. CD229 knockout mice show minimally impaired immune responses, suggesting functional redundancy with other molecules (11).
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Romero, X. et al. (2004) Tissue Antigens 64:132.
Hogarth, P.M. et al. (1980) Immunogenetics 11:65.
Mathieson, B.J. et al. (1980) J. Immunol. 125:2127.
Romero, X. et al. (2005) J. Immunol. 174:7033.
Del Valle, J.M. et al. (2003) J. Biol. Chem. 278:17430.
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Flow Cytometry: Human CD229/SLAMF3 APC‑conjugated Antibody [FAB1898A].
Peripheral blood mononuclear cells (PBMCs)
Other Experimental Details
Other Experimental Details
Gated on lymphocyte and monocyte FSC-A and SSC-A populations. Singlets were further gated on by FSC-A/FSC-H doublet exclusion. Specificity: Specific Sensitivity: Reasonably sensitive Buffer: 2% Human Serum, 0.5 mM EDTA in PBS Dilution: 1/100