Anti-Ly108/Slamf6 Antibody, clone 3E11

SLAM family member 6 (UniProt Q9ET39; also known as CD352, NK-T-B-antigen, NTB-A, Lymphocyte antigen 108) is encoded by the Slamf6 (also known as Ly108) gene (Gene ID 30925) in murine species. Ly108/Slamf6 belongs to the 7-member (SLAM/CD150, CD48/SLAMF2, Ly-9/CD229, 2B4/CD244, CD84/SLAMF5, Ly108/NTB-A/CD352, CRACC) signaling lymphocytic activation molecule family (SLAMF) of receptors broadly expressed in hematopoietic cells. 2B4 and CD48 interact with eath other, while rest of the family members act as self-ligands, where two identical molecules on the surface of two different cells can interact as receptor and ligand to each other. SLAMF members carry one or more copies of immunoreceptor tyrosine-based switch motif or ITSM (TxYxxV/I) in their cytoplasmic tails. Upon phosphorylation on the tyrosine residue, ITSM mediates the recruitment of SH2 domain-containing signaling molecules such as SLAM-associated protein (SAP), including SAP in T-, NK, and NK T-cells, EAT-2A and EAT-2B (murine) in NK cells and APCs. Ly108/Slamf6 is synthesized with a signal peptide sequence (mouse a.a. 1-30), the removal of which yields the mature Ly108 with an extracellular region (a.a. 31-239) that contains a V-type and a C2-type Ig-like domain (a.a. 36-130 and 147-210), followed by a single transmembrane domain (a.a. 240-262) and a cytoplasmic tail (a.a. 263-351). Ly108/Slamf6 cytoplasmic domain contains two copies of ITSM (mouse a.a. 293-TVYAQV-298 and 317-TIYSIV-322). Alternative splicings produce three Ly108 isoforms, Ly108-1 (Ly108s; UniProt Q9ET39-2), Ly108-2 (Ly108l; UniProt Q9ET39-1) and Ly108-3 (Not reported by UniProt), that differ in their cytoplasmic C-terminal end sequence, but are identical in the rest of their sequence.

Clone 3E11 specifically detected exogenously expressed murine Ly108-1 (Ly108s; UniProt Q9ET39-2) and Ly108-2 (Ly108l; UniProt Q9ET39-1), but not other SLAM family members (2B4, CD84, CRACC, Ly-9, and SLAM) on the surface of respective mouse BI-141 T cell transfectants. Murine Ly108 spliced isoforms Ly108-1 (Ly108s; UniProt Q9ET39-2), Ly108-2 (Ly108l; UniProt Q9ET39-1) and Ly108-3 (Not reported by UniProt) differ in their cytoplasmic C-terminal end sequence, but are identical in the rest of their sequence, including the extracellular region targeted by clone 3E11 (Zhong, M.C., and Veillette, A. (2008). J. Biol. Chem. 283(28):19255 -19264).

Epitope: extracellular domain

Murine Ly108-human IgG1 Fc fusion protein.

Anti-Anti-Ly108/Slamf6 Antibody, clone 3E11 , Cat. No. MABF919, is a highly specific rat monoclonal antibody that targets Ly108/Slamf6 and has been tested in Flow Cytometry, Function Assay, Immunoprecipitation, and Western Blotting.

Flow Cytometry Analysis: A representative lot was pre-conjugated with Alexa Fluor^™ 647 and detected surface Ly108 immunoreactivity on murine Dendritic cells (DCs), T-, B-, and NK T-cells, but not on most NK cells (Zhong, M.C., and Veillette, A. (2008). J. Biol. Chem. 283(28):19255 -19264).

Function Assay: A representative lot, together with a rabbit anti-rat IgG polyclonal antibody, cross-linked surface Ly108 on isolated C57BL/6 mouse thymocytes, resulting in an induction of SAP- and FynT-dependent Ly108 tyrosine phosphorylation (Zhong, M.C., and Veillette, A. (2008). J. Biol. Chem. 283(28):19255 -19264).

Flow Cytometry Analysis: A representative lot was pre-conjugated with Alexa Fluor 647 and detected the expression of exogenously transfected murine Ly108-1 and Ly108-2 isoforms, but not other SLAM family members (2B4, CD84, CRACC, Ly-9, and SLAM) on the surface of respective mouse BI-141 T cell transfectants (Zhong, M.C., and Veillette, A. (2008). J. Biol. Chem. 283(28):19255 -19264).

Immunoprecipitation Analysis: A representative lot, together with a rabbit anti-rat IgG polyclonal antibody, immunoprecipitated surface Ly108 from isolated C57BL/6 mouse thymocytes (Zhong, M.C., and Veillette, A. (2008). J. Biol. Chem. 283(28):19255 -19264).

Western Blotting Analysis: A representative lot detected the exogenously expressed murine Ly108-1 and Ly108-2 isoforms in lysates from respective mouse BI-141 T cell transfectants (Zhong, M.C., and Veillette, A. (2008). J. Biol. Chem. 283(28):19255 -19264).

Research Category
Inflammation & Immunology

Evaluated by Flow Cytometry analysis of mouse splenocytes.

Flow Cytometry Analysis (FC): 1 µg of this antibody detected Ly108/Slamf6 on the surface of one million mouse splenocytes.

33.26/36.44 kDa (mature/pro-Lys108-1; UniProt Q9ET39-2), 35.46/38.64 kDa (mature/pro-Lys108-2; UniProt Q9ET39-1), 35.94/39.12 kDa ((mature/pro-Lys108-3) calculated. A broad band ~65 kDa reported (Zhong, M.C., and Veillette, A. (2008). J. Biol. Chem. 283(28):19255 -19264) due to posttranslational glycosylation and phosphorylation.

Protein G purified.

Format: Purified

Purified rat monoclonal IgG2a in buffer containing PBS without preservatives.

Stable for 1 year at -20°C from date of receipt.
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.

Concentration: Please refer to lot specific datasheet.

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