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Anti-Noxa 抗体 [114C307] (ab13654)

Reviews (6)Q&A (4)References (78)
Western blot - Anti-Noxa antibody [114C307] (ab13654)
  • Flow Cytometry (Intracellular) - Anti-Noxa antibody [114C307] (ab13654)
  • Immunohistochemistry paraffin embedded sections - Anti-Noxa antibody [114C307] (ab13654)

Key features and details

  • Mouse monoclonal [114C307] to Noxa
  • Suitable for: IHC-P, WB, Flow Cyt (Intra)
  • Reacts with: Mouse, Human
  • Isotype: IgG1

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-NOXA2/p67phox antibody [EPR5065] (ab109523)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-Noxa antibody [114C307]
    Noxa 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [114C307] to Noxa

  • 由来種

    Mouse

  • アプリケーション

    適用あり: IHC-P, WB, Flow Cyt (Intra)more details

  • 種交差性

    交差種: Mouse, Human

  • 免疫原

    Fusion protein containing human Noxa.

  • ポジティブ･コントロール

    • WB: RL-7 cell lysate. Flow Cyt (Intra): Jurkat cells. IHC-P: Chronic Lymphocytic Leukemia (CLL) xenograft tissue.

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab13654の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
IHC-P
Use a concentration of 5 µg/ml.

Bouin-fixed.
WB (4)
Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 11 kDa.
Flow Cyt (Intra)
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
特記事項
IHC-P
Use a concentration of 5 µg/ml.

Bouin-fixed.
WB
Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 11 kDa.
Flow Cyt (Intra)
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ターゲット情報

  • 機能

    Promotes activation of caspases and apoptosis. Promotes mitochondrial membrane changes and efflux of apoptogenic proteins from the mitochondria. Contributes to p53/TP53-dependent apoptosis after radiation exposure. Promotes proteasomal degradation of MCL1. Competes with BAK1 for binding to MCL1 and can displace BAK1 from its binding site on MCL1 (By similarity). Competes with BIM/BCL2L11 for binding to MCL1 and can displace BIM/BCL2L11 from its binding site on MCL1.

  • 組織特異性

    Highly expressed in adult T-cell leukemia cell line.

  • 配列類似性

    Belongs to the PMAIP1 family.

  • ドメイン

    The BH3 motif is essential for pro-apoptotic activity.

  • 細胞内局在

    Mitochondrion.
  • Information by UniProt

  • 参照データベース

    • 別名

      • Adult T cell leukemia derived PMA responsive antibody
      • APR antibody
      • APR_HUMAN antibody
      • ATL-derived antibody
      • Immediate early response protein APR antibody
      • Immediate-early-response protein APR antibody
      • NOXA antibody
      • Phorbol 12 myristate 13 acetate induced protein 1 antibody
      • Phorbol-12-myristate-13-acetate-induced protein 1 antibody
      • PMA induced protein 1 antibody
      • PMA-induced protein 1 antibody
      • PMA-responsive gene antibody
      • Pmaip1 antibody
      • Protein Noxa antibody
      see all

    画像

    • Western blot - Anti-Noxa antibody [114C307] (ab13654)
      Western blot - Anti-Noxa antibody [114C307] (ab13654)

      Western blot detection of Noxa in RL-7 cell (a follicular lymphoma) lysate. A protein of approximate molecular weight of 11 kDa is detected using ab13654 at 2 µg/ml.

      Western blot detection of Noxa in RL-7 cell (a follicular lymphoma) lysate. A protein of approximate molecular weight of 11 kDa is detected using ab13654 at 2 µg/ml.
    • Flow Cytometry (Intracellular) - Anti-Noxa antibody [114C307] (ab13654)
      Flow Cytometry (Intracellular) - Anti-Noxa antibody [114C307] (ab13654)
      Overlay histogram showing Jurkat cells stained with ab13654 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab13654, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1](ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol/permeabilized in 0.1% PBS-Tween used under the same conditions.

    • Immunohistochemistry paraffin embedded sections - Anti-Noxa antibody [114C307] (ab13654)
      Immunohistochemistry paraffin embedded sections - Anti-Noxa antibody [114C307] (ab13654)

      Bouin-fixed, paraffin-embedded human tonsil tissue stained for Chronic Lymphocytic Leukemia (CLL) xenograft tissue stained for Noxa using ab13654 at 1/2000 dilution in immunohistochemical analysis.

    参考文献 (78)

    ab13654 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。

    ab13654 は 78 報の論文で使用されています。

    • Park SA  et al. Role of the SIRT1/p53 regulatory axis in oxidative stress-mediated granulosa cell apoptosis. Mol Med Rep 23:N/A (2021). PubMed: 33179088
    • Xu S  et al. Cullin-5 neddylation-mediated NOXA degradation is enhanced by PRDX1 oligomers in colorectal cancer. Cell Death Dis 12:265 (2021). PubMed: 33712558
    • Latif AL  et al. BRD4-mediated repression of p53 is a target for combination therapy in AML. Nat Commun 12:241 (2021). PubMed: 33431824
    • Ochiiwa H  et al. TAS4464, a NEDD8-activating enzyme inhibitor, activates both intrinsic and extrinsic apoptotic pathways via c-Myc-mediated regulation in acute myeloid leukemia. Oncogene 40:1217-1230 (2021). PubMed: 33420360
    • Zhu R  et al. FLT3 tyrosine kinase inhibitors synergize with BCL-2 inhibition to eliminate FLT3/ITD acute leukemia cells through BIM activation. Signal Transduct Target Ther 6:186 (2021). PubMed: 34024909
    View all Publications for this product

    レビューと Q&A

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    レビューを送る 質問を送る

    1-10 of 10 Abreviews or Q&A

    Western blot abreview for Anti-Noxa antibody [114C307]

     Average
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (SW620)
    Gel Running Conditions
    Non-reduced Denaturing (15% gel)
    Loading amount
    100000 cells
    Treatment
    Drug 1µM for 24hrs
    Specification
    SW620
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 24 2022

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Noxa antibody [114C307]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (U2OS cells)
    Permeabilization
    Yes - Tween-20
    Specification
    U2OS cells
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Acetone
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 28 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Noxa antibody [114C307]

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Human Stomach Adenocarcinoma)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 6.0 Citric Acid
    Permeabilization
    Yes - Tween-20
    Specification
    Human Stomach Adenocarcinoma
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 28 2017

    Western blot abreview for Anti-Noxa antibody [114C307]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Reduced Denaturing (18% gel)
    Sample
    Human Cell lysate - whole cell (Breast)
    Specification
    Breast
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 02 2014

    Western blot abreview for Anti-Noxa antibody [114C307]

     Average
    Abreviews
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Myeloma)
    Loading amount
    30 µg
    Specification
    Myeloma
    Treatment
    1.25 uM RITA for 6 hrs
    Gel Running Conditions
    Reduced Denaturing (12%)
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
    Read More

    Manujendra Valued Customer

    Verified customer

    投稿 Jan 07 2011

    Western blot abreview for Anti-Noxa antibody [114C307]

     Average
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HL60)
    Loading amount
    20 µg
    Specification
    HL60
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jun 26 2007

    Question

    WB with mouse B-cells
    faint band at 11 kDa, but lots of stronger non-specific bands (e.g. at 20 kDa)
    loaded 1 ug protein
    12% gel
    block w/milk
    Ab: 1/1000 overnight 4 degree
    secondary: OK

    Read More
    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 a free of charge replacement with the order number xxx with ab13687.

    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

    Question

    Dear Abcam, I need some information about this product: Anti-Noxa antibody [114C307] (ab13654). I bought this product and you suggest, in the antibody protocol, this: " This antibody gave a positive signal in Jurkat cells fixed with 80% methanol/permeabilized in 0.1% PBS-Tween used under the same conditions." Do you have any references that show the high endogenous level of Noxa in Jurkat cells, if the answer is yes can you send them to me? Many Regards,

    Read More
    Answer

    Thank you for your enquiry regarding ab13654. Noxa is highly expressed in adult T-cell leukemia cell line. Since Jurkat cells are T cell leukemia cell line, it is a good positive control for studying Noxa. Under the References section of Swiss-Prot site, you may be able to find some useful literature: http://www.uniprot.org/uniprot/Q13794 Currently, we have two specific references about this antibody listed on the product datasheet: 1) Sun Ret al. Toll-like receptor 3 (TLR3) induces apoptosis via death receptors and mitochondria by up-regulating the transactivating p63 isoform alpha (TAP63alpha). J Biol Chem 286:15918-28 (2011).Read more (PubMed: 21367858) 2) Morales AAet al. Distribution of Bim determines Mcl-1 dependence or codependence with Bcl-xL/Bcl-2 in Mcl-1-expressing myeloma cells. Blood 118:1329-39 (2011).Read more (PubMed: 21659544) If you need any specific publications regarding the expression on Noxa in Jurkat cells, I would suggest conducting a search on PubMed site: http://www.ncbi.nlm.nih.gov/pubmed/ I hope this helps and if I can assist further, please do not hesitate to contact me.

    Read More

    Question

    I am writing this e-mail to you because we have been experiencing problems with your monoclonal antibody to NOXA (Cat N°. ab13654) for several weeks now.  We have tested this antibody on PVDF and nitrocellulose membrane, with 20, 30 and 40 ug of proteins from Hela cells (in accordance with the abreviews provided on your website), but we always have several aspecific bands in the same range of size as the NOXA protein witch made the identification of the right band really difficult, even in overexpression experiments. Moreover, the detection threshold was really low, since we failed to detect the endogenous protein without a treatment with CisPlatin, despite the loading of as much as 50ug of proteins ( i.e. twice the amounts mentionned in the abreviews). And yet, this really is a problem because we aim to study variations in expression level of the endogenous protein. You will find enclosed a figure composed of two different PVDF membranes incubated with either your antibody (right part) or a monoclonal antibody to Noxa from Calbiochem. The samples used for this experiment are protein extracts coming from Hela cells transfected with a scrambled or Noxa siRNA and treated with Cisplatin to induce expression of endogenous NOXA. As you can see, the signal of the band corresponding to NOXA is really weak compared to the aspecific ones, despite the treatment with cisplatin. In addition the sizes are quite close, which could be a problem since it makes the detection of the endogenous, non-induced protein very delicate (if not impossible)with your antibody. In general we have been very satisfied with the quality of the Abcam products we have purchased and are planning purchase more. Since we have lost a considerable amount of time, money and energy with ab13654 we would like to know if Abcam would be willing to help us with a free antibody or at least discount on our next order.

    Read More
    Answer

    Thank you for contacting us.  I agree that the antibody is not working as expected, and I would be happy to offer you a free of charge replacement, credit, or refund if you have purchased the antibody in the past six months.  It would be very helpful for our quality investigation if you could please provide some additional details about your protocol: 1.)  What was your original order or PO number? 2.)  What blocking agent did you use and at what concentration? 3.)  What was the primary antibody dilution and incubation time? 4.)  What secondary antibody was used, at what concentration, and for how long? If you could please provide this additional information and let me know whether you would prefer a free of charge replacement antibody of your choice, a credit, or a refund, then I will be happy to process your request as soon as possible. 

    Read More

    Question

    LOT NUMBER 815787 ORDER NUMBER PR862521 DESCRIPTION OF THE PROBLEM Non-specific band, and high background SAMPLE Human multiple myeloma cell lines PRIMARY ANTIBODY Mouse monoclonal to Noxa (ab13654), 1-2 ug/ml, 1h at Rt or 4C O/N, wash 3 time with PBS+0,05% Tw-20, 5 minutes each DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED Beta actin was dected in the same membrane, the cells with nontreated samples should have low Noxa expression. ANTIBODY STORAGE CONDITIONS -20C SAMPLE PREPARATION Whole cell lystes, Lysis buffer with protease inhbitors, SDS sample buffer, heated at 100C 5 min before loading AMOUNT OF PROTEIN LOADED 20-30 ug ELECTROPHORESIS/GEL CONDITIONS 10%, 12% or 15% SDS-polyacrymide gel TRANSFER AND BLOCKING CONDITIONS Transfer 1 h at 100V constant, blocking with either 5% BSA or Skim milk SECONDARY ANTIBODY Cedralne lab anti-mouse IgG HRP labelled, 1:1000 HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Blokcing in Milk instead of PBS, blokcing for long time 3hrs. using high diltion of secondary antibody. ADDITIONAL NOTES We can detect the 9 KD CKS1B protein using anti-CKS1 antibody very easily with no nonspefic band, and we used fresh membrane without stripping to detect Noxa, but can not get rid of non specific bands.

    Read More
    Answer

    Thank you for your enquiry. Here are some comments and suggestions: 1. You mention using a high dilution of the secondary antibody? What dilution did you use? 2. Further dilution of the primary antibody, perhaps to 0.5ug/ml or 0.25ug/ml, will likely reduce background staining. 3. It might be helpful to see your blot. If you don't mind, please attach a JPEG formatted image of your blot to your reply. Thanks very much. I look forward to hearing from you.

    Read More

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