Anti-Legumain/Asparaginyl endopeptidase antibody Clone 6E3

Legumain (UniProt: Q99538; also known as EC: 3.4.22.34, Asparaginyl endopeptidase, AEP, Protease cysteine 1, Delta Secretase) is encoded by the LGMN (also known as PRSC1) gene (Gene ID: 5641) in human. AEP is a lysosomal endopeptidase that displays strict specificity for the hydrolysis of asparaginyl bonds and can simultaneously cleave both amyloid precursor protein (APP) and tau. It cleaves aspartyl bonds slowly, especially under acidic conditions. AEP is synthesized with a signal peptide (aa 1-17) and a propeptide (aa 324-433) that are cleaved off in the mature form. It exists a homodimer before removal of the propeptide and after autocatalytic processing exists as a monomer. In its zymogen form, the uncleaved propeptide blocks access to the active site. AEP is phosphorylated on serine 226 by serine-arginine protein kinase 2 (SRPK2), which accelerates its autocatalytic cleavage and enhances its enzymatic activity. The active form translocates to cytoplasm. In Alzheimer s disease brains AEP is reported to be highly phosphorylated. Depletion of AEP is shown to reduce amyloid peptide production and senile plaque formation in 5XFAD mouse brain that leads to restoration of synaptic activity and cognitive function. Three isoforms of AEP have been described that are produced by alternative splicing. (Ref.: Wang Z-H., et al. (2017). Mol. Cell 67(5); 812-825; Zhang, Z et al (2015). Nat. Commun. 6; 9762).

Clone 6E3 is a mouse monoclonal antibody that specifically detects Legumain (AEP).

His-tagged full-length recombinant human legumain.

Quality Control Testing

Evaluated by Western Blotting in SH-SY5Y cell lysate.

Western Blotting Analysis: A 1:1000 dilution of this antibody detected Legumain/AEP in SH-SY5Y cell lysate.

Tested Applications

Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected Legumain/AEP in mouse brain tissue lysate.

ELISA Analysis: A representative lot detected Legumain/AEP in ELISA applications (Mattock, K.L., et al. (2010). Atherosclerosis.208(1):83-9; Kang, S.S., et al. (2018). EMBO J. 37(12):e98878).

Immunoprecipitation Analysis: A representative lot immunoprecipitated legumain/AEP in Immunoprecipitation applications (Zhang, Z., et al. (2015). Nat Commun. 6:8762).

Immunohistochemistry (Paraffin) Analysis: A 1:50 dilution from a representative lot detected Legumain/AEP in human placenta tissue sections.

Immunocytochemistry Analysis: A representative lot detected Legumain/AEP in Immunocytochemistry applications (Li, D.N., et al. (2003). J. Biol. Chem. 278(40):38980-90; Wang, Z-H., et al. (2019). Cell Rep. 28(3):655-669).

Western Blotting Analysis: A representative lot detected Legumain/AEP in Western Blotting applications (Probst-Kepper, M., et al. (2009). J Cell Mol Med.;13(9B):3343-57; Zhang, Z., et al. (2015). Nat Commun.;6:8762; Wang, Z-H., et al. (2017). Mol Cell.;67(5):812-825; Zhang, Z., et al. (2017). Nat Struct Mol Biol.;24(8):632-642; Kang, S.S., et al. (2018). EMBO J.;37(12):e98878; Wang, Z-H., et al. (2018). Nat Commun.;9(1):1784; Wang, Z-H., et al. (2018). JCI Insight.;3(16):e99007; Wang, Z-H., et al. (2019). Cell Rep.;28(3):655-669).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Anti-LGMN/AEP, clone 6E3, Cat. No. MABN2304, is a mouse monoclonal antibody that detects Legumain and is tested for use in ELISA, Immunocytochemistry, Immunohistochemistry (Paraffin), Immunoprecipitation, and Western Blotting.

Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Recommend storage at +2°C to +8°C. For long term storage antibodies can be kept at -20°C. Avoid repeated freeze-thaws.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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.