ANTI-LY108/SLAMF6 CLONE 3E11

General description

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.

Specificity

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).

Immunogen

Epitope: extracellular domain

Murine Ly108-human IgG1 Fc fusion protein.

Application

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

Quality

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.

Target description

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.

Physical form

Format: Purified

Protein G purified.

Purified rat monoclonal IgG2a in buffer containing PBS without preservatives.

Storage and Stability

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.

Other Notes

Concentration: Please refer to lot specific datasheet.

Legal Information

ALEXA FLUOR is a trademark of Life Technologies

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.