Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937)

Knockout Tested Rabbit Recombinant Monoclonal BACE1 antibody. Carrier free. Suitable for IP, WB, IHC-Fr, IHC-P and reacts with Mouse, Rat, Human samples.

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Images

Western blot - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (AB238937), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	IHC-Fr	IHC-P
Human	Expected	Tested	Expected	Expected
Mouse	Tested	Expected	Tested	Tested
Rat	Tested	Expected	Expected	Tested

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes IHC-Fr is recommended for mouse only.

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info -	Notes IHC-Fr is recommended for mouse only.
Species Rat	Dilution info -	Notes IHC-Fr is recommended for mouse only.

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes IHC-P is recommended for mouse only. Binding in rat is weak under our experimental conditions and requires further optimization. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes IHC-P is recommended for mouse only. Binding in rat is weak under our experimental conditions and requires further optimization. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info -	Notes IHC-P is recommended for mouse only. Binding in rat is weak under our experimental conditions and requires further optimization. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

See full target information BACE1

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 (PubMed:10656250, PubMed:10677483, PubMed:20354142). Cleaves CHL1 (By similarity).

Alternative names

BACE, KIAA1149, BACE1, Beta-secretase 1, Aspartyl protease 2, Beta-site amyloid precursor protein cleaving enzyme 1, Memapsin-2, Membrane-associated aspartic protease 2, ASP2, Asp 2, Beta-site APP cleaving enzyme 1

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal BACE1 antibody. Carrier free. Suitable for IP, WB, IHC-Fr, IHC-P and reacts with Mouse, Rat, Human samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR19523
Purification technique: Affinity purification Protein A
Specificity: This antibody shows very low cross-reactivity with BACE2.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab238937 is the carrier-free version of Anti-BACE1 antibody [EPR19523] ab183612.

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 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 batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

BACE1 also known as beta-site APP cleaving enzyme 1 or beta-secretase plays an important role in the cleavage of amyloid precursor protein (APP). This cleavage results in the production of amyloid-beta peptides which are associated with Alzheimer's disease. BACE1 is a membrane-bound aspartic protease and has a molecular weight of approximately 50100 Da. The enzyme expresses mostly in the brain's neurons and some secretory tissues.

Biological function summary

BACE1 initiates the amyloidogenic pathway of APP processing which involves amyloid-beta generation. BACE1 doesn't function alone but acts as part of a complex that aids in protein substrate recognition and processing. Its activity contributes to physiological processes like myelination and axonal guidance indicating its importance beyond amyloid production.

Pathways

BACE1 is integral to the amyloidogenic pathway which is important in Alzheimer's disease development. It interacts with proteins such as presenilin 1 a part of the gamma-secretase complex that further processes the amyloid-beta precursor. Furthermore BACE1 links to synaptic functions and neural signaling pathways highlighting its multifaceted roles.

Associated diseases and disorders

BACE1 holds significance in Alzheimer's disease due to its role in amyloid-beta peptide production. This connection has led to the development of BACE1 inhibitors as potential therapeutic agents. Additionally BACE1’s involvement in other neural functions ties it to cognitive impairments. It also relates to APP through its function in Alzheimer's suggesting targeted strategies for treatment could involve modulating BACE1 activity.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

Western blot - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937)
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 - Anti-BACE1 antibody [EPR19523] ab183612 observed at 68 kDa. Red - loading control, Anti-GAPDH antibody [EPR16891] - Loading Control ab181602, observed at 37 kDa.
Anti-BACE1 antibody [EPR19523] 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. Anti-BACE1 antibody [EPR19523] ab183612 and Anti-GAPDH antibody [EPR16891] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-BACE1 antibody [EPR19523] ab183612).
All lanes: Western blot - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937)
Predicted band size: 56 kDa
Observed band size: 68 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937)
Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling BACE1 with Anti-BACE1 antibody [EPR19523] ab183612 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (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) (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-BACE1 antibody [EPR19523] ab183612).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937)
Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue labeling BACE1 with Anti-BACE1 antibody [EPR19523] ab183612 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-BACE1 antibody [EPR19523] ab183612).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937)
Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling BACE1 with Anti-BACE1 antibody [EPR19523] ab183612 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-BACE1 antibody [EPR19523] ab183612).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937)
BACE1 was immunoprecipitated from 1mg of rat hippocampus whole cell lysate with Anti-BACE1 antibody [EPR19523] ab183612 at 1/50 dilution. Western blot was performed from the immunoprecipitate using Anti-BACE1 antibody [EPR19523] ab183612 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: Rat hippocampus whole cell lysate, 10μg (Input).
Lane 2: Anti-BACE1 antibody [EPR19523] ab183612 IP in Rat hippocampus whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-BACE1 antibody [EPR19523] ab183612 in rat hippocampus whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-BACE1 antibody [EPR19523] ab183612).
All lanes: Immunoprecipitation - Anti-BACE1 antibody [EPR19523] (Anti-BACE1 antibody [EPR19523] ab183612)
Predicted band size: 56 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937)
Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling BACE1 with Anti-BACE1 antibody [EPR19523] ab183612 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (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 Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-BACE1 antibody [EPR19523] ab183612).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937)
BACE1 was immunoprecipitated from 1mg of mouse hippocampus whole cell lysate with Anti-BACE1 antibody [EPR19523] ab183612 at 1/50 dilution. Western blot was performed from the immunoprecipitate using Anti-BACE1 antibody [EPR19523] ab183612 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: Mouse hippocampus whole cell lysate, 10μg (Input).
Lane 2: Anti-BACE1 antibody [EPR19523] ab183612 IP in mouse hippocampus whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-BACE1 antibody [EPR19523] ab183612 in Mouse hippocampus whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-BACE1 antibody [EPR19523] ab183612).
All lanes: Immunoprecipitation - Anti-BACE1 antibody [EPR19523] (Anti-BACE1 antibody [EPR19523] ab183612)
Predicted band size: 56 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937)
Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue labelling BACE1 with Anti-BACE1 antibody [EPR19523] ab183612 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (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) (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-BACE1 antibody [EPR19523] ab183612).
Immunohistochemistry (Frozen sections) - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937)
Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse hippocampus tissue labeling BACE1 with Anti-BACE1 antibody [EPR19523] ab183612 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-BACE1 antibody [EPR19523] ab183612).
Western blot - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937)
False colour image of Western blot: Anti-BACE1 antibody [EPR19523] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red.

In Western blot, Anti-BACE1 antibody [EPR19523] ab183612 was shown to bind specifically to BACE1. A band was observed at 60/70 kDa in wild-type SH-SY5Y cell lysates with no signal observed at this size in Bace1 knockout cell line ab280078 (knockout cell lysate ab280137).

To generate this image, wild-type and Bace1 knockout SH-SY5Y cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

This data was developed using the same antibody clone in a different buffer formulation (Anti-BACE1 antibody [EPR19523] ab183612).
Lanes 1 - 3: Western blot - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937) at 1/1000 dilution
Lanes 1 - 3: Western blot - Anti-BACE1 antibody [EPR19523] (Anti-BACE1 antibody [EPR19523] ab183612) at 1/1000 dilution
Lane 1: Wild-type SH-SY5Y cell lysate at 20 µg
Lane 2: Western blot - Human BACE1 knockout SH-SY5Y cell line (ab280078)
Lane 2: Western blot - Human BACE1 knockout SH-SY5Y cell lysate (ab280137)
Lane 2: Bace1 knockout SH-SY5Y cell lysate at 20 µg
Lane 3: HAP1 cell lysate at 20 µg
Secondary
All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 60 kDa, 70 kDa
Western blot - Anti-BACE1 antibody [EPR19523] - BSA and Azide free (ab238937)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-BACE1 antibody [EPR19523] ab183612).
Blocking and dilution buffer: 5% NFDM/TBST.
Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as a loading control at 1/1000000 dilution.
We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results.
Anti-BACE1 antibody [EPR22802-233] ab263901 could be an alternative for getting stronger signal in testing cell lines.
Lanes 1 - 3: Western blot - Anti-BACE1 antibody [EPR19523] (Anti-BACE1 antibody [EPR19523] ab183612) at 1/1000 dilution
Lanes 4 - 6: Western blot - Anti-BACE1 antibody [EPR22802-233] (Anti-BACE1 antibody [EPR22802-233] ab263901) at 1/1000 dilution
Lanes 1 and 4: SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg
Lanes 2 and 5: Mouse brain tissue lysate at 20 µg
Lanes 3 and 6: Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 56 kDa
Observed band size: 68 kDa
Exposure time: 180s