Anti-BACE1 Antibody, clone 3D5

Beta-secretase 1 (UniProt: P56817; also known as EC:3.4.23.46, Aspartyl protease 2, ASP2, Asp 2, Beta-site amyloid precursor protein cleaving enzyme 1, Beta-site APP cleaving enzyme 1, Memapsin-2, Membrane-associated aspartic protease 2) is encoded by the BABACE, KIAA1149) gene (Gene ID: 23621) in human. BACE1 is a single-pass type I membrane, monomeric, glycoprotein that is synthesized with a signal peptide (aa 1-21), which is subsequently cleaved off to produce the mature protein that contains an extracellular domain (aa 22-457), a transmembrane domain (aa 458-478), and a cytoplasmic domain (aa 479-501). It also contains a DXXLL motif (aa 496-500) that is required for its proper endocytosis and retrograde transport to the trans-Golgi network, as well as for regulation of lysosomal degradation. BACE1 displays broad endopeptidase specificity and is a rate limiting enzyme that is responsible for the proteolytic processing of amyloid precursor protein (APP). It cleaves at the N-terminus of the Aβ peptide sequence, between Met671 and Asp672 of APP to yield soluble APPβ and C99. The -secretase can then cleave the C99 fragment at either Val711 or Ile713 to produce the shorter Aβ peptide (Aβ1-40) or the longer Aβ peptide (Aβ1-42). Aβ plaques are reported to induce BACE1 activity in surrounding neurons at early stages of Alzheimer s disease before neuronal death occurs. BACE1 undergoes acetylation at multiple lysine residues (126, 275, 279, 285, 299, 300, and 307) that regulates its maturation, transport to plasma membrane, and contributes to its stability. It also undergoes phosphorylation at serine 498, which is required for its interaction with GGA1 and retrograded transport from endosomal compartment to the trans-Golgi network. Six isoforms of BACE1 have been described that are produced by alternative splicing. (Ref.: Sadleir, KR., et al. (2013). Curr Alzheimer Res. 10(8); 785-789; Zhao, J., et all (2007). J. Neurosci. 27(14); :3639 -3649; Constantini, C., et al. (2007). Biochem. J. 407(3); 383-395).

His-tagged recombinant fragment corresponding to 415 amino acids from the internal region of human β-secretase 1 (BACE1).

Quality Control Testing

Evaluated by Western Blotting in SH-SY5Y cell lysate.

Western Blotting Analysis: A 1:500 dilution of this antibody detected BACE1 in SH-SY5Y cell lysate.

Tested Applications

Western Blotting Analysis: A 1:500 dilution from a representative lot detected BACE1 in NIH/3T3 cell lysate.

Immunohistochemistry Applications: A representative lot detected BACE1 in Immunohistochemistry applications (Zhao, J., et al. (2007). J Neurosci. 27(14):3639-49).

Immunofluorescence Analysis: A representative lot detected BACE1 in Immunofluorescence applications (Zhao, J., et al. (2007). J Neurosci. 27(14):3639-49).

Immunocytochemistry Analysis: A 1:25 dilution from a representative lot detected BACE1 in HeLa cells.

ELISA Analysis: A representative lot detected BACE1 in ELISA applications (Zhao, J., et al. (2007). J Neurosci. 27(14):3639-49).

Western Blotting Analysis: A representative lot detected BACE1 in Western Blotting applications (Zhao, J., et al. (2007). J Neurosci. 27(14):3639-49; Sadleir, K.R., et al. (2013). Curr Alzheimer Res. 10(8):785--9; Pigoni, M. et al. (2020). EMBO J. 39(15):e103457).

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

Clone 3D5 is a mouse monoclonal antibody that detects β-secretase 1 (BACE1).

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

0.5 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommended storage: +2°C to +8°C.

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

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