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Key Documents

MABN2640

Sigma-Aldrich

Anti-BACE1 Antibody, clone 3D5

Synonym(s):

ASP2, Asp 2, Aspartyl protease 2, Beta-secretase 1, Beta-site APP cleaving enzyme 1, Beta-site amyloid precursor protein cleaving enzyme 1, EC:3.4.23.46, Memapsin-2, Membrane-associated aspartic protease 2

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified antibody

antibody product type

primary antibodies

clone

3D5, monoclonal

mol wt

calculated mol wt 56.7 kDa
observed mol wt ~57 kDa

purified by

using protein G

species reactivity

human, mouse

packaging

antibody small pack of 100

technique(s)

ELISA: suitable
immunocytochemistry: suitable
immunofluorescence: suitable
immunohistochemistry: suitable
western blot: suitable

isotype

IgG1κ

epitope sequence

Internal

Protein ID accession no.

UniProt accession no.

storage temp.

2-8°C

Gene Information

human ... BACE1(23621)

Specificity

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

Immunogen

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

Application

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.

Target description

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).

Physical form

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

Reconstitution

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

Storage and Stability

Recommended storage: +2°C to +8°C.

Other Notes

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

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.

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Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 1

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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