Application

Flow Cytometry Analysis: A representative lot detected doxycycline-dependent expression induction of exogenously transfected CLEC-2/CLEC1B using T-REx 293 cells (Gitz, E., et al. (2014). Blood. 124(14):2262-2270).

Flow Cytometry Analysis: A representative lot (pre-conjugated with Alexa Fluor^® 488) detected CLEC-2/CLEC1B expression on the surface of platelets and CD41-positive microparticles in platelet-rich plasma (PRP), but not on monocytes, neutrophils, dendritic cells, B- or T-cells (Gitz, E., et al. (2014). Blood. 124(14):2262-2270).

Function Assay: Clone AYP1 Fab fragment (2.5 µg/mL) cross-linked with anti-mouse Fab-specific F(ab)2 fragments, but not AYP1 Fab or anti-mouse F(ab)2 alone, induced surface P-selectin expression and the aggregation of washed human platelets (Gitz, E., et al. (2014). Blood. 124(14):2262-2270).

Immunoprecipitation Analysis: A representative lot immunoprecipitated CLEC-2/CLEC1B from human platelet lysates. Rhodocytin stimulation prior to cell lysis and IP induced CLEC-2/CLEC1B tyrosine phosphorylation (Gitz, E., et al. (2014). Blood. 124(14):2262-2270).

This mouse monoclonal Anti-CLEC-2 Antibody, clone AYP1, Cat. No. MABF957 is validated for use in Flow Cytometry, Function Assay, and Immunoprecipitation for the detection of CLEC-2.

General description

C-type lectin domain family 1 member B (UniProt Q9P126; also known as C-type lectin-like receptor 2, CLEC-2) is encoded by the CLEC1B (also known as CLEC2, UNQ721/PRO1384) gene (Gene ID 51266) in human. CLEC-2 is a type II single-transmembrane (a.a. 34-54) protein with a large extracellular (a.a. 55-229) region that contains a C-type lectin domain (a.a. 109-217) and a short N-terminal cytoplasmic tail (a.a. 1-33) with a single hem-immunoreceptor tyrosine-based activation motif (hemITAM; a.a. 7-10). CLEC-2 expression is restricted to platelets, where it mediates platelets activation via its hemITAM motif, leading to proteolytic cleavage of two other platelet ITAM receptors, glycoprotein (GP)VI and Fc RIIa, but not of CLEC-2 itself. CLEC-2 functions as the receptor for the type I transmembrane GP podoplanin that is widely expressed outside of the vasculature, including lymphatic endothelial cells, type 1 lung alveolar cells, lymph node stromal cells, the choroid plexus epithelium, inflammatory macrophages, as well as a subset of activated T-helper (Th)17 cells. Patients with rheumatoid arthritis show increased plasma levels of CLEC-2-positive microparticles. These microparticles are derived from activated platelets and are negative for GPVI, while microparticles in healthy donors are predominantly derived from megakaryocytes and are positive for both CLEC-2 and GPVI. Platelet-specific deletion of CLEC-2 or one of its downstream signaling proteins, Syk, SLP-76, or PLC 2, leads to a number of developmental problems in mice, including blood-lymphatic mixing in midgestation.

Immunogen

His-tagged recombinant human CLEC-2 extracellular fragment.

Legal Information

ALEXA FLUOR is a registered trademark of Life Technologies

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Format: Purified

Quality

Evaluated by Flow Cytometry in human platelets.

Flow Cytometry Analysis: 0.2 µL of this antibody detected CLEC-2/CLEC1B on the surface of human platelets.

Specificity

Clone AYP1 targets an extracellular epitope present in both spliced isoforms of human CLEC-2/CLEC1B reported by UniProt (Q9P126).

Target description

26.60 kDa (isoform 1) and 23.10 kDa (isoform 2). ~32/40 kDa doublet reported due to differential glycosylation (Gitz, E., et al. (2014). Blood. 124(14):2262-2270).