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使用敲除细胞株进行验证重组RabMAb

重组Anti-WSTF抗体[EP1704Y] (ab51256)

Reviews (2) Submit a question References (8)
Flow Cytometry (Intracellular) - Anti-WSTF antibody [EP1704Y] (ab51256)
  • Immunocytochemistry/ Immunofluorescence - Anti-WSTF antibody [EP1704Y] (ab51256)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WSTF antibody [EP1704Y] (ab51256)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WSTF antibody [EP1704Y] (ab51256)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WSTF antibody [EP1704Y] (ab51256)
  • Western blot - Anti-WSTF antibody [EP1704Y] (ab51256)
  • Western blot - Anti-WSTF antibody [EP1704Y] (ab51256)
  • Western blot - Anti-WSTF antibody [EP1704Y] (ab51256)
  • Western blot - Anti-WSTF antibody [EP1704Y] (ab51256)
  • Anti-WSTF antibody [EP1704Y] (ab51256)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1704Y] to WSTF
  • Suitable for: IHC-P, WB, Flow Cyt (Intra), ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alkaline Phosphatase APC Carrier Free HRP PE

概述

  • 产品名称

    Anti-WSTF抗体[EP1704Y]
    参阅全部 WSTF 一抗

  • 描述

    兔单克隆抗体[EP1704Y] to WSTF

  • 宿主

    Rabbit

  • 经测试应用

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

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide within Human WSTF (C terminal). The exact sequence is proprietary.

  • 阳性对照

    • WB: Wild-type HAP1, HeLa (ab150035), and PC12 cell lysates. Mouse and Rat testis. B16-F0, MCF7, HeLa whole cell lysates. Flow cyt: HeLa cells; IHC: rat cardiac muscle tissue, mouse cardiac muscle tissue, human breast carcinoma; ICC/IF: HeLa cells.

  • 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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab51256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
1/700. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB (2)
1/15000 - 1/20000. Detects a band of approximately 185 kDa (predicted molecular weight: 171 kDa).
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF
1/50.

For unpurified format use at 1/100 to 1/250 dilution
说明
IHC-P
1/700. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
1/15000 - 1/20000. Detects a band of approximately 185 kDa (predicted molecular weight: 171 kDa).
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF
1/50.

For unpurified format use at 1/100 to 1/250 dilution

靶标

  • 功能

    Atypical tyrosine-protein kinase that plays a central role in chromatin remodeling and acts as a transcription regulator. Involved in DNA damage response by phosphorylating 'Tyr-142' of histone H2AX (H2AXY142ph). H2AXY142ph plays a central role in DNA repair and acts as a mark that distinguishes between apoptotic and repair responses to genotoxic stress. Essential component of the WICH complex, a chromatin remodeling complex that mobilizes nucleosomes and reconfigures irregular chromatin to a regular nucleosomal array structure. The WICH complex regulates the transcription of various genes, has a role in RNA polymerase I and RNA polymerase III transcription, mediates the histone H2AX phosphorylation at 'Tyr-142', and is involved in the maintenance of chromatin structures during DNA replication processes. In the complex, it mediates the recruitment of the WICH complex to replication foci during DNA replication. Also involved in vitamin D-coupled transcription regulation via its association with the WINAC complex, a chromatin-remodeling complex recruited by vitamin D receptor (VDR), which is required for the ligand-bound VDR-mediated transrepression of the CYP27B1 gene. In the WINAC complex, plays an essential role by targeting the complex to acetylated histones, an essential step for VDR-promoter association.

  • 组织特异性

    Ubiquitously expressed with high levels of expression in heart, brain, placenta, skeletal muscle and ovary.

  • 疾病相关

    Note=BAZ1B is located in the Williams-Beuren syndrome (WBS) critical region. WBS results from a hemizygous deletion of several genes on chromosome 7q11.23, thought to arise as a consequence of unequal crossing over between highly homologous low-copy repeat sequences flanking the deleted region. Haploinsufficiency of BAZ1B may be the cause of certain cardiovascular and musculo-skeletal abnormalities observed in the disease.

  • 序列相似性

    Belongs to the WAL family. BAZ1B subfamily.
    Contains 1 bromo domain.
    Contains 1 DDT domain.
    Contains 1 PHD-type zinc finger.
    Contains 1 WAC domain.

  • 发展阶段

    Expressed at equal levels in 19-23 weeks old fetal tissues.

  • 结构域

    The N-terminal part (1-345), including the WAC domain and the C motif, mediates the tyrosine-protein kinase activity.
    The bromo domain mediates the specific interaction with acetylated histones.

  • 翻译后修饰

    Phosphorylated upon DNA damage, probably by ATM or ATR.

  • 细胞定位

    Nucleus. Accumulates in pericentromeric heterochromatin during replication. Targeted to replication foci throughout S phase via its association with PCNA.
  • Information by UniProt

  • 数据库链接

    • 别名

      • baz1b antibody
      • BAZ1B_HUMAN antibody
      • Bromodomain adjacent to zinc finger domain protein 1B antibody
      • hWALP 2 antibody
      • hWALP-2 antibody
      • hWALP2 antibody
      • transcription factor WSTF antibody
      • Tyrosine-protein kinase BAZ1B antibody
      • WALP-2 antibody
      • WALP2 antibody
      • WBRS 9 antibody
      • WBRS-9 antibody
      • WBRS9 antibody
      • WBSC 10 antibody
      • WBSC-10 antibody
      • WBSC10 antibody
      • WBSCR10 antibody
      • WBSCR9 antibody
      • Williams Beuren syndrome chromosome region 9 protein antibody
      • Williams syndrome transcription factor antibody
      • Williams-Beuren syndrome chromosomal region 10 protein antibody
      • Williams-Beuren syndrome chromosomal region 9 protein antibody
      • WSTF antibody
      see all

    图片

    • Flow Cytometry (Intracellular) - Anti-WSTF antibody [EP1704Y] (ab51256)
      Flow Cytometry (Intracellular) - Anti-WSTF antibody [EP1704Y] (ab51256)
      Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling WSTF with Purified ab51256 at 1:20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
    • Immunocytochemistry/ Immunofluorescence - Anti-WSTF antibody [EP1704Y] (ab51256)
      Immunocytochemistry/ Immunofluorescence - Anti-WSTF antibody [EP1704Y] (ab51256)
      Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling WSTF with Purified ab51256 at 1:50 dilution (2.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WSTF antibody [EP1704Y] (ab51256)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WSTF antibody [EP1704Y] (ab51256)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cardiac muscle tissue sections labeling WSTF with Purified ab51256 at 1:700 dilution (0.211 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WSTF antibody [EP1704Y] (ab51256)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WSTF antibody [EP1704Y] (ab51256)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cardiac muscle tissue sections labeling WSTF with Purified ab51256 at 1:700 dilution (0.211 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WSTF antibody [EP1704Y] (ab51256)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WSTF antibody [EP1704Y] (ab51256)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling WSTF with Purified ab51256 at 1:700 dilution (0.211 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
    • Western blot - Anti-WSTF antibody [EP1704Y] (ab51256)
      Western blot - Anti-WSTF antibody [EP1704Y] (ab51256)
      All lanes : Anti-WSTF antibody [EP1704Y] (ab51256) at 1/5000 dilution

      Lane 1 : B16-F0 (Mouse melanoma epithelial cell-like) whole cell lysate
      Lane 2 : Mouse testis lysate
      Lane 3 : Rat testis lysate

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 171 kDa
      Observed band size: 185 kDa why is the actual band size different from the predicted?



      Exposure time

      Lane 1: 10 seconds

      Lane 2: 40 seconds

      Lane 3: 180 seconds

    • Western blot - Anti-WSTF antibody [EP1704Y] (ab51256)
      Western blot - Anti-WSTF antibody [EP1704Y] (ab51256)
      All lanes : Anti-WSTF antibody [EP1704Y] (ab51256) at 1/15000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : BAZ1B knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 171 kDa
      Observed band size: 175 kDa why is the actual band size different from the predicted?



      Lanes 1- 2: Merged signal (red and green). Green - ab51256 observed at 175 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

       ab51256 was shown to react with WSTF in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264907 (knockout cell lysate ab257370) was used. Wild-type HeLa and WSTF knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab51256 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 15000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-WSTF antibody [EP1704Y] (ab51256)
      Western blot - Anti-WSTF antibody [EP1704Y] (ab51256)
      All lanes : Anti-WSTF antibody [EP1704Y] (ab51256) at 1/20000 dilution

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate
      Lane 3 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 171 kDa



      Exposure time:

      Left image: 180 seconds

      Right image: 40 seconds

      We recommend to use ab109439 for WSTF Western Blot testing because ab51256 detects nonspecific bands.

    • Western blot - Anti-WSTF antibody [EP1704Y] (ab51256)
      Western blot - Anti-WSTF antibody [EP1704Y] (ab51256)
      All lanes : Anti-WSTF antibody [EP1704Y] (ab51256) at 1/15000 dilution

      Lane 1 : Wild-type HAP1 cell lysate
      Lane 2 : WSTF knockout HAP1 cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : PC12 cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 171 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab51256 observed at 175 kDa. Red - loading control, ab18058, observed at 124 kDa.

      ab51265 was shown to recognize WSTF in wilt-type cells along with additional cross-reactive bands as signal was lost in WSTF knockout samples. Wild-type and WSTF knockout samples were subjected to SDS-PAGE. ab51256 and ab18058 (loading control to vinculin) were diluted 1/15 000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Anti-WSTF antibody [EP1704Y] (ab51256)
      Anti-WSTF antibody [EP1704Y] (ab51256)

    实验方案

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    文献 (8)

    发表研究结果有使用 ab51256？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab51256 被引用在 8 文献中.

    • D'Alesio C  et al. RNAi screens identify CHD4 as an essential gene in breast cancer growth. Oncotarget 7:80901-80915 (2016). WB . PubMed: 27779108
    • Sun H  et al. BAZ1B in Nucleus Accumbens Regulates Reward-Related Behaviors in Response to Distinct Emotional Stimuli. J Neurosci 36:3954-61 (2016). PubMed: 27053203
    • Liu Y  et al. KRASG12 mutant induces the release of the WSTF/NRG3 complex, and contributes to an oncogenic paracrine signaling pathway. Oncotarget 7:53153-53164 (2016). PubMed: 27449290
    • Ashton NW  et al. Novel insight into the composition of human single-stranded DNA-binding protein 1 (hSSB1)-containing protein complexes. BMC Mol Biol 17:24 (2016). WB . PubMed: 27938330
    • Sun H  et al. ACF chromatin-remodeling complex mediates stress-induced depressive-like behavior. Nat Med 21:1146-53 (2015). WB . PubMed: 26390241
    • Vidi PA  et al. NuMA promotes homologous recombination repair by regulating the accumulation of the ISWI ATPase SNF2h at DNA breaks. Nucleic Acids Res 42:6365-79 (2014). ICC/IF ; Human . PubMed: 24753406
    • Sánchez-Molina S  et al. Role for hACF1 in the G2/M damage checkpoint. Nucleic Acids Res 39:8445-56 (2011). WB . PubMed: 21745822
    • Yoshimura K  et al. Distinct function of 2 chromatin remodeling complexes that share a common subunit, Williams syndrome transcription factor (WSTF). Proc Natl Acad Sci U S A 106:9280-5 (2009). ICC/IF ; Mouse . PubMed: 19470456

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    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-WSTF antibody [EP1704Y]

     Average
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Vascular smooth muscle cells)
    Gel Running Conditions
    Reduced Denaturing (6%)
    Loading amount
    30 µg
    Specification
    Vascular smooth muscle cells
    Blocking step
    Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 Feb 13 2017

    Western blot abreview for Anti-WSTF antibody [EP1704Y]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Non-reduced Denaturing (12% SDS-PAGE)
    Sample
    Human Cell lysate - nuclear (HeLa)
    Specification
    HeLa
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 Sep 05 2013

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