重组Anti-BACE1抗体[EPR19523] (ab183612)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19523] to BACE1
- Suitable for: IHC-P, WB, IP, IHC-Fr
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-BACE1抗体[EPR19523]
参阅全部 BACE1 一抗 -
描述
兔单克隆抗体[EPR19523] to BACE1 -
宿主
Rabbit -
经测试应用
适用于: IHC-P, WB, IP, IHC-Frmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- 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.
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常规说明
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
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. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR19523 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab183612于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
IHC-P | (8) |
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) |
1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 56 kDa).
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IP |
1/50.
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IHC-Fr |
1/250.
IHC-Fr is recommended for mouse only. |
说明 |
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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. |
靶标
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功能
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. -
组织特异性
Expressed at high levels in the brain and pancreas. In the brain, expression is highest in the substantia nigra, locus coruleus and medulla oblongata. -
序列相似性
Belongs to the peptidase A1 family. -
结构域
The transmembrane domain is necessary for its activity. It determines its late Golgi localization and access to its substrate, APP. -
翻译后修饰
Glycosylated. -
细胞定位
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. - Information by UniProt
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数据库链接
- 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
see all -
别名
- APP beta secretase antibody
- Asp 2 antibody
- ASP2 antibody
see all
图片
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All lanes : Anti-BACE1 antibody [EPR19523] (ab183612) at 1/1000 dilution
Lane 1 : Wild-type SH-SY5Y cell lysate
Lane 2 : Bace1 knockout SH-SY5Y cell lysate
Lane 3 : HAP1 cell lysate
Lysates/proteins at 20 µg per lane.
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.
Predicted band size: 56 kDa
Observed band size: 60,70 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-BACE1 antibody [EPR19523] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red.
In Western blot, 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.
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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.
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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.
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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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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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 secondsAn 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.
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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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
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.
-
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.
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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.
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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 Detection Reagent (HRP) (ab131366), was used for detection 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.
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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 Detection Reagent (HRP) (ab131366), was used for detection 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.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (28)
ab183612 被引用在 28 文献中.
- Liu L et al. Hypercholesterolemia aggravates sevoflurane-induced cognitive impairment in aged rats by inducing neurological inflammation and apoptosis. J Biochem Mol Toxicol 36:e23009 (2022). PubMed: 35174938
- Yang Y et al. Ginsenoside Rg1 improves Alzheimer's disease by regulating oxidative stress, apoptosis, and neuroinflammation through Wnt/GSK-3β/β-catenin signaling pathway. Chem Biol Drug Des 99:884-896 (2022). PubMed: 35313087
- Chocron ES et al. Genetic and pharmacologic proteasome augmentation ameliorates Alzheimer's-like pathology in mouse and fly APP overexpression models. Sci Adv 8:eabk2252 (2022). PubMed: 35675410
- Wang Y et al. Krüppel-like factor 5 accelerates the pathogenesis of Alzheimer's disease via BACE1-mediated APP processing. Alzheimers Res Ther 14:103 (2022). PubMed: 35883144
- Wang X et al. TRPV1 Modulator Ameliorates Alzheimer-Like Amyloid-β Neuropathology via Akt/Gsk3β-Mediated Nrf2 Activation in the Neuro-2a/APP Cell Model. Oxid Med Cell Longev 2022:1544244 (2022). PubMed: 36065437