MABF957
Anti-CLEC-2 Antibody, clone AYP1
clone AYP1, from mouse
Synonym(s):
C-type lectin domain family 1 member B, C-type lectin-like receptor 2, CLEC-2
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.43
Recommended Products
biological source
mouse
Quality Level
antibody form
purified immunoglobulin
antibody product type
primary antibodies
clone
AYP1, monoclonal
species reactivity
human
technique(s)
flow cytometry: suitable
isotype
IgG1κ
NCBI accession no.
UniProt accession no.
shipped in
ambient
target post-translational modification
unmodified
Gene Information
human ... CLEC1B(51266)
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.
Specificity
Clone AYP1 targets an extracellular epitope present in both spliced isoforms of human CLEC-2/CLEC1B reported by UniProt (Q9P126).
Immunogen
His-tagged recombinant human CLEC-2 extracellular fragment.
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).
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.
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.
Flow Cytometry Analysis: 0.2 µL of this antibody detected CLEC-2/CLEC1B on the surface of human platelets.
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).
Physical form
Format: Purified
Other Notes
Concentration: Please refer to lot specific datasheet.
Legal Information
ALEXA FLUOR is a trademark of Life Technologies
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Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 2
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Manisha Yadav et al.
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