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  1. Link

    apaf1-antibody-ab2000.pdf

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Cell Biology Apoptosis Mitochondrial
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Anti-APAF1 抗体 (ab2000)

  • Datasheet
Reviews (2)Q&A (4)References (7)

Product price, shipping and contact information

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出荷および受注について

Abpromise

保証された製品品質、優れたカスタマー・サポート。 詳細はこちら。

Western blot - Anti-APAF1 antibody (ab2000)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APAF1 antibody (ab2000)

Key features and details

  • Rabbit polyclonal to APAF1
  • Suitable for: IHC-P, WB
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

こちらの製品もご検討ください

タンパク質
Product image
Recombinant Human APAF1 protein (Tagged) (ab198064)
二次抗体
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

関連製品

製品の概要

  • 製品名

    Anti-APAF1 antibody
    APAF1 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to APAF1
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: IHC-P, WBmore details
  • 種交差性

    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide corresponding to Human APAF1 aa 1158-1177 (C terminal).
    Sequence:

    TFYTNGTNLKKIHVSPDFKT


    (Peptide available as ab7872)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 特記事項

    Apoptosis Protease Activating Factor

    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.

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • バッファー

    pH: 7.2
    Preservative: 0.02% Sodium azide
  • Concentration information loading...
  • 精製度

    Immunogen affinity purified
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Cell Biology
    • Apoptosis
    • Mitochondrial
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Cytochrome C
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Cytochrome C
    • Cancer
    • Cell Death
    • Apoptosis
    • Mitochondrial
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism

関連製品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human APAF1 protein (Tagged) (ab198064)

アプリケーション

Our Abpromise guarantee covers the use of ab2000 in the following tested applications.

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

アプリケーション Abreviews 特記事項
IHC-P Use a concentration of 20 µg/ml.
WB 1/500 - 1/2000. Predicted molecular weight: 130 kDa.

ターゲット情報

  • 機能

    Oligomeric Apaf-1 mediates the cytochrome c-dependent autocatalytic activation of pro-caspase-9 (Apaf-3), leading to the activation of caspase-3 and apoptosis. This activation requires ATP. Isoform 6 is less effective in inducing apoptosis.
  • 組織特異性

    Ubiquitous. Highest levels of expression in adult spleen and peripheral blood leukocytes, and in fetal brain, kidney and lung. Isoform 1 is expressed in heart, kidney and liver.
  • 配列類似性

    Contains 1 CARD domain.
    Contains 1 NB-ARC domain.
    Contains 13 WD repeats.
  • ドメイン

    The CARD domain mediates interaction with APIP.
  • 細胞内局在

    Cytoplasm.
  • Target information above from: UniProt accession O14727 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 参照データベース

    • Entrez Gene: 317 Human
    • Entrez Gene: 11783 Mouse
    • Entrez Gene: 78963 Rat
    • Omim: 602233 Human
    • SwissProt: O14727 Human
    • SwissProt: O88879 Mouse
    • SwissProt: Q9EPV5 Rat
    • Unigene: 552567 Human
    • Unigene: 728891 Human
    • Unigene: 220289 Mouse
    • Unigene: 64522 Rat
    see all
  • 別名

    • APAF 1 antibody
    • Apaf-1 antibody
    • APAF_HUMAN antibody
    • Apaf1 antibody
    • Apoptotic peptidase activating factor 1 antibody
    • Apoptotic protease activating factor 1 antibody
    • Apoptotic protease activating factor antibody
    • Apoptotic protease-activating factor 1 antibody
    • CED 4 antibody
    • CED4 antibody
    • KIAA0413 antibody
    see all

画像

  • Western blot - Anti-APAF1 antibody (ab2000)
    Western blot - Anti-APAF1 antibody (ab2000)
    All lanes : Anti-APAF1 antibody (ab2000) at 1 µg/ml

    Lane 1 : Human heart tissue lysate with absence of blocking peptide
    Lane 2 : Human heart tissue lysate with presence of blocking peptide

    Predicted band size: 130 kDa
    Observed band size: 130 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APAF1 antibody (ab2000)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APAF1 antibody (ab2000)
    Immunohistochemistry of Apaf1 in human skin tissue with Apaf1 antibody at 20 µg/ml.

プロトコール

  • Immunohistochemistry protocols
  • Western blot protocols

Click here to view the general protocols

データシートおよび資料

    • Datasheet
  • 参考文献 (7)

    ab2000 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab2000 は 7 報の論文で使用されています。

    • Zhang X  et al. The effects of umbilical cord-derived macrophage exosomes loaded with cisplatin on the growth and drug resistance of ovarian cancer cells. Drug Dev Ind Pharm N/A:1-13 (2020). PubMed: 32482115
    • Pan Z  et al. Cinobufagin Induces Cell Cycle Arrest at the G2/M Phase and Promotes Apoptosis in Malignant Melanoma Cells. Front Oncol 9:853 (2019). PubMed: 31552178
    • Wang SQ  et al. Knockdown of IGF-1R Triggers Viral RNA Sensor MDA5- and RIG-I-Mediated Mitochondrial Apoptosis in Colonic Cancer Cells. Mol Ther Nucleic Acids 16:105-117 (2019). PubMed: 30861413
    • Liu Y  et al. Breviscapine ameliorates CCl4-induced liver injury in mice through inhibiting inflammatory apoptotic response and ROS generation. Int J Mol Med 42:755-768 (2018). PubMed: 29717768
    • Gulic T  et al. Granulysin-mediated apoptosis of trophoblasts in blighted ovum and missed abortion. Am J Reprod Immunol 80:e12978 (2018). PubMed: 29774968
    • Hathaway QA  et al. Maternal-engineered nanomaterial exposure disrupts progeny cardiac function and bioenergetics. Am J Physiol Heart Circ Physiol 312:H446-H458 (2017). PubMed: 28011589
    • Nichols CE  et al. Cardiac and mitochondrial dysfunction following acute pulmonary exposure to mountaintop removal mining particulate matter. Am J Physiol Heart Circ Physiol 309:H2017-30 (2015). PubMed: 26497962

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-6 of 6 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-APAF1 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Lymph node)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Ventana CC2 (Citrate)
    Permeabilization
    No
    Specification
    Lymph node
    Fixative
    Paraformaldehyde
    Read More

    Dr. Li Li

    Verified customer

    投稿 Jun 05 2017

    Question

    The Abcam Order # is**********.

    For ab32351 and ab2000 customer would like to receive ab59388 and ab2001 as replacements.

    For ab93942she agreed to submit an Abreview, I asked her to create an account on Abcam's website and submitthe Abreview. She said she is already doing it.

    Thank you and Regards,

    Read More

    Abcam community

    Verified customer

    Asked on Feb 09 2012

    Answer

    Thank you for your reply.

    I have processed your request to receive the 2 free of charge replacements and below is the testing discount code for ab93942.

    Please let me know if there is anything else I can help you with.

    DISCOUNT CODE:**********
    Expiration date: *********

    I am very pleased to hear you would like to accept our offer and test ab93942 in rat tissue. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview forrat tissue and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


    Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

    Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

    The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

    Read More

    Abcam Scientific Support

    Answered on Feb 09 2012

    Question

    Customer said the secondary ab6721is working fine with other Abcam antibodies they bought in the same order, all from rabbit:

    - Beta actin = ab8227
    - Synaptophysin = ab52636
    - OPA1 = ab101589

    Read More

    Abcam community

    Verified customer

    Asked on Jan 24 2012

    Answer

    Thank you for your reply.

    I am sorry that the customer has had problems with these antibodies. As we guarantee thatboth ab2000 and ab32351 to work in western blot on rat tissue and they are not working as we expect then I am happy to either replace or refund the cost of these antibodies. We have multiple other antibodies against these targets and I would be happy to send you any of those.

    As ab93942 was only predicted to work in rat, then it does not fall under our Abpromise, but what I can do is offer them a back dated testing code. They would have to submit anAbreview on how the antibody performed and once that Abreview was submitted, they would be able to use the testing discount code to get another antibody free of charge.

    Please let me know how you would like to proceed and I will be happy to resolve this issue for you.

    Read More

    Abcam Scientific Support

    Answered on Jan 24 2012

    Question

    For the Sci Sup,

    Customer is getting no bands with 3 different antibodies (ab93942, ab32351 and ab2000). Please, find below the questionnaire filled out by the customer. Let me know if you need further information.

    Abcam WB Questionnaire



    1) Abcam product code:



    ab93942 (Rabbit polyclonal to DNM1L)

    ab32351 (Rabbit monoclonal [E87] to Caspase 3)

    ab2000 (Rabbit polyclonal to APAF1)



    2) Lot number:



    3) Antibody storage conditions (temperature/reconstitution etc):



    All antibodies at -20ºC.



    4) Description of the problem (high background, wrong band size, more bands, no band etc.):



    We can’t see bands for all antibodies.



    5) Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.):



    Wistar rats’ brain (hippocampus, cortex and striatum).



    6) Sample preparation (Buffer/Protease inhibitors/Heating sample etc.):



    For the first time, the extraction was made with RIPA buffer with protease inhibitor.

    After, we tried a different extraction (for mitochondrial and cytosolic proteins → 10mM Tris-HCl pH 8, 1mM EDTA pH 8, 100mM NaCl, protease inhibitor cocktail, 0,5% Triton X-100 and 0,1% SDS).



    7) Amount of protein loaded:



    For the first time we tried 25ug of protein. After, we performed a protein curve, using 20, 40 and 50ug of protein. We also tried to boil the sample to facilitate the binding of antibodies.



    8) Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.):



    Acrylamide gel 10% and 12%, SDS-page, electrophoresis for 2h / 2h30 at 80 / 100v.



    9) Transfer and blocking conditions (Buffer/time period, Blocking agent etc.):



    Transfer was made on ice for 2h at 350mA; 2h at 50mA; 2h30 at 85mA; 2h30 at 100mA; 3h at 100mA. Membranes were blocked with skim milk 5% for 15 minutes or albumin 5% for 30 minutes.



    10) Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step):



    DNM1L (ab93942) = 1:500.

    Caspase 3 (ab32351) = 1:5000, 1:3000, 1:1500, 1:1000, 1:500.

    APAF1 (ab2000) = 1:1000 and 1:500. All antibodies were incubated overnight, on ice, shaking. After, the membranes were washed with TBST 1X (6x for 5 minutes).



    11) Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step):



    The secondary antibody used was Goat pAb to Rb IgG (HRP) – ab6721. The concentration was 1:2000 for 1h on ice, shaking. After, the membranes were washed with TBST 1X (6x for 5 minutes).



    12) Detection method (ECL, ECLPlus etc.):



    First, we used Western Lightning ECL kit – Perkin Elmer. 500uL and 1000uL of each reagent and exposure for 1, 2, 3, 10 and 15 minutes.

    We also used 500uL and 1000uL of ECL Western Blotting Substrate Kit (ab65628) for others tests and exposure for 15 seconds, 30 seconds, 1 minute, 1.5 minutes and 3 minutes.



    13) Positive and negative controls used (please specify):



    We used marker all the time and used an optimized antibody to check if our sample was ok.



    14) How many times have you tried the Western?



    For DNM1L we tried 5 times. For caspase 3 we tried 7 times. For APAF1 we tried 10 times.



    15) Have you run a "No Primary" control?



    No.



    16) Do you obtain the same results every time? e.g. are the background bands always in the same place?



    Yes. All the time we can’t see bands.



    17) What steps have you altered to try and optimize the use of this antibody?



    We changed all the steps of Western Blotting. We prepared new samples with two extraction methods, we changed the gel preparation, we changed the electrophoresis and transfer buffer, we changed blocking agent. We also checked if our samples in electrophoresis and transfer were ok, using Coomassie blue dying to visualize the proteins. We also tried different antibody dilutions as well as protein loading concentrations.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 16 2012

    Answer

    Thank you for contacting Abcam.

    I am sorry about the problems that you have been having in western blot, with some of our products. Based on the information you have provided, I suspect that the problem is with the secondary antibody, since it would be really bad luck if you had all 3 primary antibodies fail to work, although for ab93942, we only predict that it will work in rat tissue, as we have not tested it rat yet.

    I would be happy to send you a new secondary antibody to see if that resolves the issue. Also, under our Abpromise, both ab2000 and ab32351 is guaranteed to work in western blot on rat tissue, therefore if it does turn out that the primary antibody is at fault, then I would be happy to replace or refund the cost of these antibodies.

    If you would like me to replace the secondary antibody, could you please provide me with the original order information for this product and also let me know if there is any particular secondary that you would like to receive.

    I look forward to your reply and helping you to resolve this issue.

    Read More

    Abcam Scientific Support

    Answered on Jan 16 2012

    Western blot abreview for Anti-APAF1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (C57Bl/6 mouse liver)
    Loading amount
    10 µg
    Specification
    C57Bl/6 mouse liver
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    投稿 Oct 19 2011

    Question

    You sale two different antibodies for apaf-1. Could you explain the difference? They are CT and NT. Please send 800 number also.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 23 2001

    Answer

    Anti-Apaf-1 NT (2013) and anti-Apaf-1 CT (2015) were gernerated with peptides selected from the N-terminus and C-terminus of the protein. 888 513 9525

    Read More

    Abcam Scientific Support

    Answered on Jan 26 2001

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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