Anti-BACE1抗体(ab2077)

Reviews (4)Q&A (11)References (65)

Key features and details

  • Rabbit polyclonal to BACE1
  • Suitable for: WB, ICC/IF, IHC-P
  • Reacts with: Mouse, Human
  • Isotype: IgG

概述

  • 产品名称

    Anti-BACE1抗体
    参阅全部 BACE1 一抗

  • 描述

    兔多克隆抗体to BACE1

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, ICC/IF, IHC-Pmore details

  • 种属反应性

    与反应: Mouse, Human

  • 免疫原

    Synthetic peptide corresponding to Human BACE1 (C terminal). The immunogen is located within the last 50 amino acids of BACE1 and it consists of a 17 aa peptide.
    Database link: P56817
    (Peptide available as ab7883)

  • 阳性对照

    • IHC-P: Mouse brain tissue WB: A431, A549, Caco-2, Daudi, HeLa, K562, Jurkat, SK-N-SH, THP-1. Human brain tissue lysate, 3T3 lysate. ICC/IF: 3T3 cells.

  • 常规说明

    Beta-site APP Cleaving Enzyme.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

性能

应用

Our Abpromise guarantee covers the use of ab2077 in the following tested applications.

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

应用 Ab评论 说明
WB Use a concentration of 1 µg/ml. Detects a band of approximately 70 kDa. We recommend overnight incubation at 4C and using 5% skim milk to block.
ICC/IF Use a concentration of 10 µg/ml.
IHC-P Use a concentration of 2.5 µg/ml.

靶标

  • 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

  • 数据库链接

    • 别名

      • 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

    图片

    • Western blot - Anti-BACE1 antibody (ab2077)
      Western blot - Anti-BACE1 antibody (ab2077)
      All lanes :

      Lane 1 : A431 (Human epidermoid carcinoma cell line) whole cell lysate with Anti-BACE1 antibody (ab2077)
      Lane 2 : A549 (Human lung carcinoma cell line) whole cell lysate with Anti-BACE1 antibody (ab2077)
      Lane 3 : Caco-2 (Human colorectal adenocarcinoma cell line) whole cell lysate with Anti-BACE1 antibody (ab2077)
      Lane 4 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysate with Anti-BACE1 antibody (ab2077)
      Lane 5 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with Anti-BACE1 antibody (ab2077)
      Lane 6 : K562 (Human chronic myelogenous leukemia cell line from bone marrow ) whole cell lysate with Anti-BACE1 antibody (ab2077)
      Lane 7 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate with Anti-BACE1 antibody (ab2077)
      Lane 8 : SK-N-SH (Human neuroblastoma cell line) whole cell lysate with Anti-BACE1 antibody (ab2077)
      Lane 9 : THP-1 (Human monocytic leukemia cell line) whole cell lysate with Anti-BACE1 antibody (ab2077)

      Lysates/proteins at 15 µg per lane.

      Blocking peptides at 1 µg/ml per lane.

      Secondary
      All lanes : Rabbit IgG antibody (HRP) at 1/10000 dilution

      Developed using the ECL technique.

      Additional bands at: 65 kDa (possible glycosylated form)



      10% gel.

      Running conditions: 130v for 2 hours.

      Transfer conditions: wet, 250mA, 2 hrs (Nitrocellulose membrane).

      Blocking condition: 5% non-fat dry milk in TBS, 4C, overnight.

      Primary antibody incubation: Room temperature for 1 hour.

      Secondary antibody incubation: Room temperature for 1 hour. 

      Washing conditions: 15 mL TSBT, 3 x 10 minutes. 

      Exposure: ECL solution

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE1 antibody (ab2077)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BACE1 antibody (ab2077)

      Immunohistochemical analysis of paraffin-embedded mouse brain tissue using ab2077 at 2.5 µg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

    • Immunocytochemistry/ Immunofluorescence - Anti-BACE1 antibody (ab2077)
      Immunocytochemistry/ Immunofluorescence - Anti-BACE1 antibody (ab2077)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryo fibroblast cell line) cells labeling BACE1 with ab2077 at 20 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue). Image showing both membrane and cytosol staining on NIH/3T3 cells.

    • Immunocytochemistry/ Immunofluorescence - Anti-BACE1 antibody (ab2077)
      Immunocytochemistry/ Immunofluorescence - Anti-BACE1 antibody (ab2077)
      Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 (Mouse embryo fibroblast cell line) cells labeling BACE1 with ab2077 at 10 μg/mL. Cells were fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4oC. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

       

    • Western blot - Anti-BACE1 antibody (ab2077)
      Western blot - Anti-BACE1 antibody (ab2077)
      All lanes : Anti-BACE1 antibody (ab2077) at 1 µg/ml

      Lane 1 : Human brain tissue lysate with absence of blocking peptide
      Lane 2 : Human brain tissue
      lysate with BACE1 peptide (ab7883)
      Lane 3 : Mouse 3T3/NIH cell lysate

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit IgG HRP conjugate at 1/10000 dilution

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



      Incubate the antibody for 1 hour at room temperature in 5% NFDM/TBST.

       

       

    • Immunocytochemistry/ Immunofluorescence - Anti-BACE1 antibody (ab2077)
      Immunocytochemistry/ Immunofluorescence - Anti-BACE1 antibody (ab2077)Image courtesy of an anonymous Abreview.

      Immunocytochemiscal analysis of D54MG (human glioblastoma cell line) cells labeling BACE1 with ab2077.Cells were fixed in paraformaldehyde, permeabilized with 0.1% Triton X-100, blocked with 0.5% BSA for 20 minutes at room temperature, then incubated with ab2077 at a 1/50 dilution for 16 hours at 4°C. The secondary used was a TRITC conjugated goat anti-rabbit polyclonal, used at a 1/400 dilution. Nuclei are counterstained with DAPI.

      See Abreview

    文献 (65)

    发表研究结果有使用 ab2077？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab2077 被引用在 65 文献中.

    • Ye JY  et al. ß-Sitosterol treatment attenuates cognitive deficits and prevents amyloid plaque deposition in amyloid protein precursor/presenilin 1 mice. Korean J Physiol Pharmacol 24:39-46 (2020). PubMed: 31908573
    • Tam JM  et al. A Novel Dual Fluorochrome Near-Infrared Imaging Probe for Potential Alzheimer's Enzyme Biomarkers-BACE1 and Cathepsin D. Molecules 25:N/A (2020). PubMed: 31936569
    • Chiritoiu M  et al. EDEM1 Drives Misfolded Protein Degradation via ERAD and Exploits ER-Phagy as Back-Up Mechanism When ERAD Is Impaired. Int J Mol Sci 21:N/A (2020). PubMed: 32423001
    • Long Z  et al. Dynamic changes of autophagic flux induced by Abeta in the brain of postmortem Alzheimer's disease patients, animal models and cell models. Aging (Albany NY) 12:10912-10930 (2020). PubMed: 32535554
    • Wu Q  et al. SRC-1 Knockout Exerts No Effect on Amyloid ß Deposition in APP/PS1 Mice. Front Aging Neurosci 12:145 (2020). PubMed: 32625077
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-10 of 15 Abreviews or Q&A

    Question

    Thank you for your response.

    Our customer has also requested for a credit note for this one too.

    Appreciate your time and if you could kindly organise this for us.

    Thanks again and enjoy your day J

    Read More
    Answer

    Thank you for contacting us.

    Your credit note ID is xxxxx.

    I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

    (1) Redeemed against the original invoice if this hasn't already been paid.
    (2) Held on the account for use against a future order.
    (3) A full refund can be offered where no other invoices are outstanding.

    Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

    To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

    The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

    Read More

    Question

    Thank you for your email.

    Our customer has responded with the following;

    Could you please confirm if a loading control was included and what the results were? A loading control assures that the samples used are of good quality.

    My positive control were SH-SY5Y and mice brain lysate (30ug/well). As you know both are neurons and have plenty amount of BACE1.

    2. We have some data that suggest that the BACE1 protein is difficult to transfer to the membrane in WB application. I would appreciate if you could confirm if the transfer of protein onto the membrane was assessed and what was the result? I have been working on BACE1 for more than 3 years, and I think this question is not true and accepteptable.

    3. Could you confirm the isotype that is recognised by the secondary antibody and weather it is known to work well with other primary antibodies? It was goat Anti-Rab IgG (Total) HRP

    Our all secondary antibody have been already optimized and they are working very well with 1:3000 dilution.

    I look forward to your response.

    Read More
    Answer

    Thank you for your message and for providing this further information.

    I am sorry to confirm that we can not make any further suggestions in this case to improve the results. It seems that you regrettably received a bad vial. For that reason I would be pleased to arrange a refund, credit note or free of charge replacement in compensation.

    I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Question

    Product code: 2077
    Lot number: GR1723-1
    Inquiry: Hi Technical, We have a customer who is experiencing problems with this antibody. Please find attached the customer's questionnaire.

    Read More
    Answer

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from our Anti-BACE1 antibody (ab2077) .

    The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

    I would like to reassure you that ab2077 is tested and covered by our 6 month guarantee for use in WB and mouse as well as human samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

    Regrettably reviewing all the details there are not many suggestions we can make to improve the protocol. However, I would like to investigate the case a bit further for you. Before deciding how to proceed with this particular case, I would appreciate if you can confirm some details of the procedure.

    1. Could you please confirm if a loading control was included and what the results were? A loading control assures that the samples used are of good quality.

    2. We have some data that suggest that the BACE1 protein is difficult to transfer to the membrane in WB application. I would appreciate if you could confirm if the transfer of protein onto the membrane was assased and what was the result?

    3. Could you confirm the isotype that is recognised by the secondary antibody and weather it is known to work well with other primary antibodies?

    Thank you very much for your cooperation. Please do not hesitate to contact me again with the further requested details. I look forward to your answer and am sure we will be able to find a solution.

    Read More

    Question

    The ab2077 I have was giving the expected signal for my past several lysates. To verify the exact band, since I was getting one at approx 70 and another at approx 55/60, I used the mouse lysate. The strong band at 100 kD for the mouse lysate was unexpected and confusing, leading me to believe the mouse lysate wasn't good. I thought running it again with the mouse lysate in the middle of the gel might clarify where the positive control was appearing. I also thought that even if I ran this again with a new vial of ab2077, I still wouldn't have a positive control to verify my band (and appease future reviewers).

    Having said that, perhaps you're right that I should at least try another ab2077. I'd rather do that then try a different primary antibody. So, go ahead and send a new ab2077. If I get the same thing, then I'll have to decide how to proceed to verify that the bands I'm seeing are indeed the glycosylated and non-glycosylated forms for BACE1.

    Read More
    Answer

    Thank you for your reply.

    I am sending you a new vial of ab2077, your new order number is ** and you should have the new antibody by 10.30am tomorrow morning. If there is anything else I can help you with, or if you have any issues with the new vial of the antibody please let me know.

    I hope your experiments go well.

    Read More

    Question

    I understand that you can't send all the reagents, but I've been working on this project for too long to quit. I don't think anything's wrong with the ab2077 antibody. Would you be able to send me only the 3T3 lysate? I need 30 ul to run at least 2 lanes. If that works, then I'll order (and pay for) more ab2077 to run the experiment to completion.

    Read More
    Answer

    Thank you for your reply.

    If sending you a new aliquot of the cell lysate would help you to complete your experiments, then I would be happy to do so. However, I am confused as to why you think that the current vial of ab2077 that you have is ok when it is not giving the expected signal. I was hoping that if I send you a different antibody against the same target, then you would be able to detect your band of interest.

    As I mentioned previously, I would be happy to send a new vial of cell lysate, I just wanted to confirm with you the course of action that you wish to take and to double check that you did not want to try a different primary antibody.

    I look forward to your reply.

    Read More

    Question

    Thanks for your response. For some reason the email I sent a couple days ago bounced back. Please see attached an overlay of the ECL image with the Marker image. It definitely seems like there are two bands (between 75 kD and 50 kD) that correspond with the BACE1 protein. Specifically, one at approx 70 kD and one at approx 56 kD. Based on the data sheets you sent me, the 70 kD band is probably the glycosylated form and the smaller approx 56 band might be the unglycosylated form. It is strange that the mouse lysate appears in the 100 kD area, but there seems to be a faint band in the 3T3 lanes at approx 56 kD. I need to re-run this in the middle of a gel to be sure, but I need more lysate.

    I'm not ready to jump ship and go with a different company. Is there another positive control you'd recommend? Also, since the peptide hasn't worked, I'm guessing the batch was somehow bad. I also think I should try a new BACE1 antibody along with the ab2077 I still have. So, can you please send me the following: 1) peptide ab7883, 2) either a different positive control or more of the mouse lysate 3T3 (at least 40ul), 3) ab108394 (since perhaps a monoclonal Ab would decrease the non-specific bands I'm seeing).

    Read More
    Answer

    Thank you for your reply.

    I am sorry about the issues you have been having with ab2077 in western blot. I cannot send you all of the reagents that you are requesting, I can either send 1 free antibody as a replacement for ab2077, or I can process a refund for the cost of the original purchase.

    Please let me know how you would like to proceed.

    Read More

    Question

    Not seeing the correct size band in wetern blotiing

    Read More
    Answer

    Thank you for calling Abcam yesterday.

    I am sorry about the issues you have been having with ab2077 in western blot. Having talked to my colleagues, I think the best way to move forward with this is to either replace the antibody that you currently have or to refund the cost of the original purchase.

    I do not think that there are any protocol tips that I could give that would help resolve the banding pattern that you are seeing and can only assume that the vial of ab2077 that you received is not working correctly.

    A few alternative BACE1 antibodies to consider are:

    https://www.abcam.com/BACE1-antibody-EPR3956-ab108394.html
    https://www.abcam.com/BACE1-antibody-ab63954.html



    If either of these antibodies would work for you (or if there is another BACE1 antibody that you would like to try), please let me know and I will be happy to send it out to you.

    I look forward to your reply.

    Read More

    Question

    Called because product was giving non specific bands

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    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued the free of charge blocking peptide for this product with the order number I have also included our protocol for blocking with the immunizing peptide for your review. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

    Read More

    Immunocytochemistry/ Immunofluorescence abreview for Anti-BACE1 antibody

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (murine glioma cell line C6)
    Specification
    murine glioma cell line C6
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0,1% Triton X 100
    Blocking step
    BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    提交于 Aug 30 2011

    Immunocytochemistry/ Immunofluorescence abreview for Anti-BACE1 antibody

     Average
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (human glioblastoma cell line D54MG)
    Specification
    human glioblastoma cell line D54MG
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0,1% Triton X 100
    Blocking step
    BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    提交于 Aug 30 2011

    1-10 of 15 Abreviews or Q&A

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