Anti-BACE1 antibody [EPR19523] KO Tested Recombinant (ab183612)

Product name

Anti-BACE1 antibody [EPR19523]
See all BACE1 primary antibodies
Description

Rabbit monoclonal [EPR19523] to BACE1
Host species

Rabbit
Tested applications

Suitable for: IHC-P, WB, IP, IHC-Frmore details
Species reactivity

Reacts with: Mouse, Rat
Immunogen

Recombinant fragment within Mouse BACE1 aa 50-350. The exact sequence is proprietary.
Database link: P56818

Run BLAST with Run BLAST with
Positive control
- WB: Mouse and rat hippocampus and brain lysates. IHC-P: Mouse hippocampus and cerebrum tissues; rat cerebrum tissue. IHC-Fr: Mouse hippocampus tissue. IP: Rat and mouse hippocampus whole cell lysates.
General notes

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

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19523
Isotype

IgG
Research areas

Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Immunohistochemistry kits
- EXPOSE Rabbit specific HRP/DAB detection IHC Kit (ab80437)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

Our Abpromise guarantee covers the use of ab183612 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
IHC-P		1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. IHC-P is recommended for mouse only. Binding in rat is weak under our experimental conditions and requires further optimization.
WB		1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 56 kDa).
IP		1/50.
IHC-Fr		1/250. IHC-Fr is recommended for mouse only.

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: 23821 Mouse
- Entrez Gene: 29392 Rat
- SwissProt: P56818 Mouse
- SwissProt: P56819 Rat
- Unigene: 24044 Mouse
- Unigene: 207201 Rat
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

Western blot - Anti-BACE1 antibody [EPR19523] (ab183612)

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)

Lanes 1 - 3: Merged signal (red and green). Green - ab183612 observed at 68 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab183612 was shown to specifically react with BACE1 in wild-type HAP1 cells as signal was lost in BACE1 knockout cells. Wild-type and BACE1 knockout samples were subjected to SDS-PAGE. ab183612 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 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 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-BACE1 antibody [EPR19523] (ab183612)

All lanes : Anti-BACE1 antibody [EPR19523] (ab183612) at 1/1000 dilution

Lane 1 : Mouse hippocampus lysate
Lane 2 : Rat brain lysate
Lane 3 : Rat hippocampus lysate
Lane 4 : Mouse ovary lysate
Lane 5 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate
Lane 6 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 7 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 56 kDa
Observed band size: 68 kDa (why is the actual band size different from the predicted?)

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1 and 2: 1 minute; Lane 3,4,5,6,7 and 8: 3 minutes.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE1 antibody [EPR19523] (ab183612)

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling BACE1 with ab183612 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on some neurons of the rat cerebrum is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Binding in rat was weak under our experimental conditions and requires further optimization.
Western blot - Anti-BACE1 antibody [EPR19523] (ab183612)

Anti-BACE1 antibody [EPR19523] (ab183612) at 1/1000 dilution + Mouse brain lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 56 kDa
Observed band size: 68 kDa (why is the actual band size different from the predicted?)

Exposure time: 30 seconds

An additional band was observed at 70 kD. The expression profile is consistent with what has been described in the literature (PMID: 22741101).

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE1 antibody [EPR19523] (ab183612)

Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue labeling BACE1 with ab183612 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on mouse Hilar region of the dentate gyrus is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE1 antibody [EPR19523] (ab183612)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling BACE1 with ab183612 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on neurons of the mouse cerebrum is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE1 antibody [EPR19523] (ab183612)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling BACE1 with ab183612 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on mouse liver. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE1 antibody [EPR19523] (ab183612)

Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue labelling BACE1 with ab183612 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. The sample was counterstained with hematoxylin. Antigen retrieval was performed using Tris/EDTA buffer; pH 9.0.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Binding in rat was weak under our experimental conditions and requires further optimization.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE1 antibody [EPR19523] (ab183612)

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labelling BACE1 with ab183612 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. The sample was counterstained with hematoxylin. Antigen retrieval was performed using Tris/EDTA buffer; pH 9.0.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Binding in rat was weak under our experimental conditions and requires further optimization.
Immunohistochemistry (Frozen sections) - Anti-BACE1 antibody [EPR19523] (ab183612)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse hippocampus tissue labeling BACE1 with ab183612 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). The result showed mainly cytoplasmic staining on mouse hippocampus. The nuclear counterstain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Immunoprecipitation - Anti-BACE1 antibody [EPR19523] (ab183612)

BACE1 was immunoprecipitated from 1mg of rat hippocampus whole cell lysate with ab183612 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab183612 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.

Lane 1: Rat hippocampus whole cell lysate, 10µg (Input).

Lane 2: ab183612 IP in Rat hippocampus whole cell lysate.

Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab183612 in rat hippocampus whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 5 seconds.
Immunoprecipitation - Anti-BACE1 antibody [EPR19523] (ab183612)

BACE1 was immunoprecipitated from 1mg of mouse hippocampus whole cell lysate with ab183612 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab183612 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.

Lane 1: Mouse hippocampus whole cell lysate, 10µg (Input).

Lane 2: ab183612 IP in mouse hippocampus whole cell lysate.

Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab183612 in Mouse hippocampus whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 1 second.

Immunoprecipitation protocols
Immunohistochemistry protocols
Western blot protocols

This product has been referenced in:

Cai H et al. Neuroprotective effects of bajijiasu against cognitive impairment induced by amyloid-ß in APP/PS1 mice. Oncotarget 8:92621-92634 (2017). Read more (PubMed: 29190943) »

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Anti-BACE1 antibody [EPR19523] (ab183612)

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Customer reviews and Q&As

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BACE1 antibody [EPR19523]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BACE1 antibody [EPR19523]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BACE1 antibody [EPR19523]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BACE1 antibody [EPR19523]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BACE1 antibody [EPR19523]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BACE1 antibody [EPR19523]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BACE1 antibody [EPR19523]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BACE1 antibody [EPR19523]