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  1. Link

    bace1-antibody-epr3956-ab108394.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-BACE1 antibody [EPR3956] (ab108394)

  • Datasheet
  • SDS
Reviews (2)Q&A (2)References (44)

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Western blot - Anti-BACE1 antibody [EPR3956] (ab108394)
  • Western blot - Anti-BACE1 antibody [EPR3956] (ab108394)
  • Western blot - Anti-BACE1 antibody [EPR3956] (ab108394)
  • Immunoprecipitation - Anti-BACE1 antibody [EPR3956] (ab108394)
  • Western blot - Anti-BACE1 antibody [EPR3956] (ab108394)
  • Anti-BACE1 antibody [EPR3956] (ab108394)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3956] to BACE1
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-BACE1 antibody [EPR3956]
    See all BACE1 primary antibodies
  • Description

    Rabbit monoclonal [EPR3956] to BACE1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: SH-SY5Y cell lysate and human fetal, mouse and rat brain tissue lysates. IP: Human fetal brain tissue lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, PBS, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3956
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Proteases
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteolytic enzymes
    • Aspartic protease
    • BACEs
    • Neuroscience
    • Diseases

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-BACE1 antibody [EPR19523] (ab225307)
    • Alexa Fluor® 488 Anti-BACE1 antibody [EPR19523] (ab225308)
    • Anti-BACE1 antibody [EPR3956] - BSA and Azide free (ab237595)
  • Assay kits

    • Beta Secretase Activity Assay Kit (Fluorometric) (ab65357)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Rat brain normal tissue lysate - membrane extract (ab29473)
    • Mouse brain tissue lysate - total protein (ab30151)
    • Mouse brain tissue lysate - total protein (0 days) (ab7188)
    • Mouse brain tissue lysate - total protein (14 days) (ab7189)
    • Mouse brain tissue lysate - total protein (7 days) (ab7190)
  • Recombinant Protein

    • Recombinant human BACE1 protein (Active) (ab168075)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108394 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB (2)
1/1000 - 1/10000. Predicted molecular weight: 56 kDa.
IP
1/10 - 1/100.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 56 kDa.
IP
1/10 - 1/100.
Application notes
Is unsuitable for Flow Cyt,ICC/IF or IHC-P.

Target

  • Function

    Responsible for the proteolytic processing of the amyloid precursor protein (APP). Cleaves at the N-terminus of the A-beta peptide sequence, between residues 671 and 672 of APP, leads to the generation and extracellular release of beta-cleaved soluble APP, and a corresponding cell-associated C-terminal fragment which is later released by gamma-secretase.
  • Tissue specificity

    Expressed at high levels in the brain and pancreas. In the brain, expression is highest in the substantia nigra, locus coruleus and medulla oblongata.
  • Sequence similarities

    Belongs to the peptidase A1 family.
  • Domain

    The transmembrane domain is necessary for its activity. It determines its late Golgi localization and access to its substrate, APP.
  • Post-translational
    modifications

    Glycosylated.
  • Cellular localization

    Membrane. Golgi apparatus > trans-Golgi network. Endoplasmic reticulum. Endosome. Cell surface. Predominantly localized to the later Golgi/trans-Golgi network (TGN) and minimally detectable in the early Golgi compartments. A small portion is also found in the endoplasmic reticulum, endosomes and on the cell surface.
  • Target information above from: UniProt accession P56817 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 23621 Human
    • Entrez Gene: 23821 Mouse
    • Entrez Gene: 29392 Rat
    • Omim: 604252 Human
    • SwissProt: P56817 Human
    • SwissProt: P56818 Mouse
    • SwissProt: P56819 Rat
    • Unigene: 504003 Human
    • Unigene: 24044 Mouse
    • Unigene: 207201 Rat
    see all
  • Alternative names

    • APP beta secretase antibody
    • Asp 2 antibody
    • ASP2 antibody
    • Aspartyl protease 2 antibody
    • BACE 1 antibody
    • BACE antibody
    • BACE1 antibody
    • BACE1_HUMAN antibody
    • Beta secretase 1 antibody
    • Beta secretase antibody
    • Beta site amyloid beta A4 precursor protein cleaving enzyme antibody
    • Beta site amyloid precursor protein cleaving enzyme 1 antibody
    • Beta site amyloid precursor protein cleaving enzyme antibody
    • Beta site APP cleaving enzyme 1 antibody
    • Beta site APP cleaving enzyme antibody
    • Beta-secretase 1 antibody
    • Beta-site amyloid precursor protein cleaving enzyme 1 antibody
    • Beta-site APP cleaving enzyme 1 antibody
    • FLJ90568 antibody
    • HSPC104 antibody
    • Memapsin 2 antibody
    • Memapsin-2 antibody
    • Memapsin2 antibody
    • Membrane associated aspartic protease 2 antibody
    • Membrane-associated aspartic protease 2 antibody
    • Transmembrane aspartic proteinase Asp2 antibody
    see all

Images

  • Western blot - Anti-BACE1 antibody [EPR3956] (ab108394)
    Western blot - Anti-BACE1 antibody [EPR3956] (ab108394)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: BACE1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: SHSY5Y whole cell lysate (20 µg)
    Lane 4: Human brain whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab108394 observed at 70 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab108394 was shown to recognize BACE1 in wild type cells as signal was lost at the expected MW in BACE1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and BACE1 knockout samples were subjected to SDS-PAGE. Ab108394 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-BACE1 antibody [EPR3956] (ab108394)
    Western blot - Anti-BACE1 antibody [EPR3956] (ab108394)
    All lanes : Anti-BACE1 antibody [EPR3956] (ab108394) at 1/10000 dilution (purified)

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size: 56 kDa
    Observed band size: 70 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-BACE1 antibody [EPR3956] (ab108394)
    Western blot - Anti-BACE1 antibody [EPR3956] (ab108394)
    Anti-BACE1 antibody [EPR3956] (ab108394) at 1/10000 dilution (purified) + Human fetal brain tissue lysate at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size: 56 kDa
    Observed band size: 70 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-BACE1 antibody [EPR3956] (ab108394)
    Immunoprecipitation - Anti-BACE1 antibody [EPR3956] (ab108394)

    ab108394 (purified) at 1/40 immunoprecipitating BACE1 in human fetal brain whole tissue lysate.

    Lane 1 (input): human fetal brain whole tissue lysate (10µg)

    Lane 2 (+): ab108394 + human fetal brain whole tissue lysate (10µg).

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab108394 in human fetal brain whole tissue lysate.

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-BACE1 antibody [EPR3956] (ab108394)
    Western blot - Anti-BACE1 antibody [EPR3956] (ab108394)
    All lanes : Anti-BACE1 antibody [EPR3956] (ab108394) at 1/1000 dilution (unpurified)

    Lane 1 : Fetal brain lysate
    Lane 2 : SH-SY5Y lysate
    Lane 3 : Mouse brain lysate
    Lane 4 : Rat brain lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 56 kDa

  • Anti-BACE1 antibody [EPR3956] (ab108394)
    Anti-BACE1 antibody [EPR3956] (ab108394)

Protocols

  • Western blot protocols
  • Immunoprecipitation protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (44)

Publishing research using ab108394? Please let us know so that we can cite the reference in this datasheet.

ab108394 has been referenced in 44 publications.

  • Pi T  et al. High Methionine Diet-Induced Alzheimer's Disease like Symptoms Are Accompanied by 5-Methylcytosine Elevated Levels in the Brain. Behav Neurol 2021:6683318 (2021). PubMed: 33880134
  • Liu Y  et al. Plasmalogen attenuates the development of hepatic steatosis and cognitive deficit through mechanism involving p75NTR inhibition. Redox Biol 43:102002 (2021). PubMed: 33984602
  • Langness VF  et al. Cholesterol-lowering drugs reduce APP processing to Aß by inducing APP dimerization. Mol Biol Cell 32:247-259 (2021). PubMed: 33296223
  • Maitra S  et al. Phenanthroline impairs ßAPP processing and expression, increases p53 protein levels and induces cell cycle arrest in human neuroblastoma cells. Brain Res Bull 170:29-38 (2021). PubMed: 33556560
  • Zhong M  et al. Nasal Delivery of D-Penicillamine Hydrogel Upregulates a Disintegrin and Metalloprotease 10 Expression via Melatonin Receptor 1 in Alzheimer's Disease Models. Front Aging Neurosci 13:660249 (2021). PubMed: 33935689
View all Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Western blot abreview for Anti-BACE1 antibody [EPR3956]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Hippocampus)
Gel Running Conditions
Non-reduced Denaturing
Loading amount
40 µg
Specification
Hippocampus
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Hyeon Joo Ham

Verified customer

Submitted Dec 28 2018

Western blot abreview for Anti-BACE1 antibody [EPR3956]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Brain)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (8%)
Loading amount
30 µg
Treatment
250 μg/kg LPS for 1 week
Specification
Brain
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Ms. Ji Hye Han

Verified customer

Submitted Jul 05 2018

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