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Anti-XAF1抗体(ab81353)

Submit a review Q&A (2)References (4)
Western blot - Anti-XAF1 antibody (ab81353)
  • Immunocytochemistry/ Immunofluorescence - Anti-XAF1 antibody (ab81353)

Key features and details

  • Rabbit polyclonal to XAF1
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF
  • Reacts with: Human
  • Isotype: IgG

概述

  • 产品名称

    Anti-XAF1抗体
    参阅全部 XAF1 一抗

  • 描述

    兔多克隆抗体to XAF1

  • 宿主

    Rabbit

  • 经测试应用

    适用于: Flow Cyt (Intra), WB, ICC/IFmore details

  • 种属反应性

    与反应: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: HepG2 cell lysate ICC: MCF7 cells

  • 常规说明

    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

性能

  • 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.

  • 存储溶液

    Constituents: 76.92% Trehalose, 17.31% Sodium chloride, 3.85% Disodium hydrogenorthophosphate
  • Concentration information loading...

  • 纯度

    Immunogen affinity purified

  • 克隆

    多克隆

  • 同种型

    IgG

  • 研究领域

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab81353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
Flow Cyt (Intra)
Use 1-3µg for 106 cells.
WB
Use a concentration of 0.1 - 0.5 µg/ml. Predicted molecular weight: 35 kDa.
ICC/IF
Use a concentration of 5 µg/ml.
说明
Flow Cyt (Intra)
Use 1-3µg for 106 cells.
WB
Use a concentration of 0.1 - 0.5 µg/ml. Predicted molecular weight: 35 kDa.
ICC/IF
Use a concentration of 5 µg/ml.

靶标

  • 功能

    Seems to function as a negative regulator of members of the IAP (inhibitor of apoptosis protein) family. Inhibits anti-caspase activity of BIRC4. Induces cleavage and inactivation of BIRC4 independent of caspase activation. Mediates TNF-alpha-induced apoptosis and is involved in apoptosis in trophoblast cells. May inhibit BIRC4 indirectly by activating the mitochondrial apoptosis pathway. After translocation to mitochondra, promotes translocation of BAX to mitochondria and cytochrome c release from mitochondria. Seems to promote the redistribution of BIRC4 from the cytoplasm to the nucleus, probably independent of BIRC4 inactivation which seems to occur in the cytoplasm. The BIRC4-XAF1 complex mediates down-regulation of BIRC5/survivin; the process requires the E3 ligase activity of BIRC4. Seems to be involved in cellular sensitivity to the proapoptotic actions of TRAIL. May be a tumor suppressor by mediating apoptosis resistance of cancer cells.

  • 组织特异性

    Widely expressed. Expression is frequently down-regulated in cancer cell lines. Isoform 5 is widely expressed. Expressed in placenta (at protein level).

  • 序列相似性

    Contains 1 TRAF-type zinc finger.

  • 发展阶段

    Selectively expressed in third trimester placenta.

  • 细胞定位

    Nucleus and Cytoplasm. Nucleus. Mitochondrion. Found in the cytoplasm and nucleus of placental syncytiotrophoblasts. Translocates to mitochondria upon TNF-alpha treatment.
  • Information by UniProt

  • 数据库链接

    • 别名

      • BIRC 4 binding protein antibody
      • BIRC4 binding protein antibody
      • BIRC4-binding protein antibody
      • BIRC4BP antibody
      • HSXIAPAF 1 antibody
      • HSXIAPAF1 antibody
      • XAF 1 antibody
      • XAF1 antibody
      • XAF1_HUMAN antibody
      • XIAP associated factor 1 antibody
      • XIAP-associated factor 1 antibody
      • XIAPAF1 antibody
      see all

    图片

    • Western blot - Anti-XAF1 antibody (ab81353)
      Western blot - Anti-XAF1 antibody (ab81353)
      All lanes :

      All lanes : HEPG2 whole cell lysate

      Lysates/proteins at 40 µg per lane.

      Predicted band size: 35 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-XAF1 antibody (ab81353)
      Immunocytochemistry/ Immunofluorescence - Anti-XAF1 antibody (ab81353)
      ICC/IF image of ab81353 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab81353, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h.Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    文献 (4)

    发表研究结果有使用 ab81353？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab81353 被引用在 4 文献中.

    • Zhu C  et al. SARM1 deficiency up-regulates XAF1, promotes neuronal apoptosis, and accelerates prion disease. J Exp Med 216:743-756 (2019). PubMed: 30842236
    • Park GB  et al. Ampelopsin-induced reactive oxygen species enhance the apoptosis of colon cancer cells by activating endoplasmic reticulum stress-mediated AMPK/MAPK/XAF1 signaling. Oncol Lett 14:7947-7956 (2017). PubMed: 29250183
    • Li K  et al. Adenovirus encoding XAF-1 and TNF-a in the same open reading frame efficiently inhibits hepatocellular cancer cells. Mol Med Rep 13:5169-76 (2016). PubMed: 27121136
    • Liu RT  et al. Inflammatory mediators induced by amyloid-beta in the retina and RPE in vivo: implications for inflammasome activation in age-related macular degeneration. Invest Ophthalmol Vis Sci 54:2225-37 (2013). PubMed: 23462752

    客户评价及客户问答

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    1-2 of 2 Abreviews or Q&A

    Question

    LOT NUMBER XXXX ORDER NUMBER XXXX DESCRIPTION OF THE PROBLEM Non-specific band Can not detect endogenous and exogenous human XAF-1. SAMPLE Transiently transfected HEK293 cells. Whole cell lysate PRIMARY ANTIBODY Concentration or dilution : 1:400 Diluent buffer : 5% non-fat milk TBST Incubation time : O/N Incubation temperature: 4C Wash Buffer : TBST( 0.1% Tween-20) Number of washes : 3times DETECTION METHOD Enhanced luminol reagent POSITIVE AND NEGATIVE CONTROLS USED Positive control : Exogenous Flag tagged human XAF-1 Negative control : X ANTIBODY STORAGE CONDITIONS -20C SAMPLE PREPARATION Lysis buffer : 1% triton x-100 in PBS buffer. Protease inhibitors: roche protease inhibitor cocktail. Phosphatase inhibitors : X Reducing agent : X Boiling for ≥5 min? yes AMOUNT OF PROTEIN LOADED 60ug/lane ELECTROPHORESIS/GEL CONDITIONS 10.5% TRANSFER AND BLOCKING CONDITIONS Type of membrane : PVDF Protein transfer verified : 2hr, 90V Blocking agent and concentration : 5% non-fat milk Blocking time : 1hr Blocking temperature : R/T SECONDARY ANTIBODY Species: Goat Isotype: IgG-HRP Reacts against: Rb Concentration or dilution : 1:3000 Diluent buffer : 5% non-fat milk TBST Incubation time : 1hr Incubation temperature: R/T Fluorochrome or enzyme conjugate: HRP Wash Buffer : TBST( 0.1% Tween-20) Number of washes : 3times HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? No WHAT STEPS HAVE YOU ALTERED? - ADDITIONAL NOTES -

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    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 this antibody.

    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.

    Reviewing this case, I would like to offer some suggestions to help optimise the results from ab81353.

    This protein presents multiple isoforms at different molecular weights. In such cases sample preparation is crucial. I would recommend you to lysate your samples using RIPA buffer and using phosphatase as well as protease inhibitors. This is crucial to avoid protein degradation.
    We also recommend using reducing agents unless otherwise stated on the datasheet. This will ensure the protein is in the correct conformation to run at the correct Molecular Weight and for the antibody to run. Mercaptoethanol and DTT are commonly used reducing agents.
    Reduce protein load from 60ug to 20ug. The non-specific bands could be due to too much sample loaded.
    Increase the blocking conditions. Try blocking for two hours instead of only one, still at RT.
    In cases where multiple non specific bands are present, the use of a positive control becomes indispensable to help identify the target protein.
    In order to dismiss non specific bands coming from the secondary, I would run a non primary control, incubating only with the secondary. If this were the case, I would then suggest decreasing the secondary antibody concentration, to 1/5000.



    I hope these tips help, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again and I’ll be more than happy to help you further.

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    Question

    Hi, I intend to study the expression of XAF-1 in breast cancer tissues using immunohistochemistry. I have both ab17204 and ab81353. After looking at the images on IHC posted in the datasheets of these 2 antibodies, I would like to know if the team has carried out antigen retrieval (if yes, which method, using which buffer?) during the IHC on those tissues which gave you the pictures in the datasheet. Thank you for your assistance. Best regards,

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    Answer

    Thank you for your enquiry. I can confirm the following antigen retrieval methods were used in testing of these antibodies. Please not that this may require individual optimization by customers in their own experiments: ab17204 XAF1 antibody We use a citrate buffer, pH 5.5 and heat for antigen retrieval. ab81353 XAF1 antibody Heat repair: Immerse the paraffin sections into Citrate buffer (AR0024, pH6.0), heat until boiling in the microwave and then cut off the power, keep it in the microwave for nearly 5~10 mins. Repeat 1 or 2 times. Then cool at room temperature and wash 1 or 2 times with PBS (pH7.2-7.6). I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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