Anti-WHSC1/NSD2抗体[29D1] - ChIP Grade (ab75359)

概述

  • 产品名称Anti-WHSC1/NSD2抗体[29D1] - ChIP Grade
    参阅全部 WHSC1/NSD2 一抗
  • 描述
    小鼠单克隆抗体[29D1] to WHSC1/NSD2 - ChIP Grade
  • 经测试应用适用于: CHIPseq, WB, IP, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    Fusion protein, corresponding to amino acids 1-647 of Human GST-WHSC1/NSD2

  • 阳性对照
    • This antibody gave a positive result in IHC in the following FFPE tissue: Human normal testis. In Western Blot, this antibody gave a positive signal in HeLa and SHSY5Y whole cell lysates.

应用

Our Abpromise guarantee covers the use of ab75359 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
CHIPseq Use at an assay dependent concentration. PubMed: 22196736
WB Use a concentration of 5 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 152 kDa).
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

靶标

  • 功能Probable histone methyltransferase (By similarity). May act as a transcription regulator that binds DNA and suppresses IL5 transcription.
  • 组织特异性Widely expressed.
  • 疾病相关Note=A chromosomal aberration involving WHSC1 is a cause of multiple myeloma tumors. Translocation t(4;14)(p16.3;q32.3) with IgH.
    Note=WHSC1 is located in the Wolf-Hirschhorn syndrome (WHS) critical region. WHS results from by sub-telomeric deletions in the short arm of chromosome 4. WHSC1 is deleted in every case, however deletion of linked genes contributes to both the severity of the core characteristics and the presence of the additional syndromic problems.
  • 序列相似性Belongs to the histone-lysine methyltransferase family. SET2 subfamily.
    Contains 1 AWS domain.
    Contains 1 HMG box DNA-binding domain.
    Contains 4 PHD-type zinc fingers.
    Contains 1 post-SET domain.
    Contains 2 PWWP domains.
    Contains 1 SET domain.
  • 翻译后修饰Phosphorylated upon DNA damage, probably by ATM or ATR.
  • 细胞定位Cytoplasm and Nucleus. Chromosome.
  • Information by UniProt
  • 数据库链接
  • 别名
    • FLJ23286 antibody
    • IL5 promoter REII region binding protein antibody
    • KIAA1090 antibody
    • MGC176638 antibody
    • MMSET antibody
    • MMSET type II antibody
    • Multiple myeloma SET domain containing protein type III antibody
    • Multiple myeloma SET domain protein antibody
    • Multiple myeloma SET domain-containing protein antibody
    • NSD 2 antibody
    • NSD2 antibody
    • NSD2_HUMAN antibody
    • Nuclear SET domain-containing protein 2 antibody
    • Probable histone-lysine N-methyltransferase NSD2 antibody
    • Protein trithorax-5 antibody
    • REIIBP antibody
    • Trithorax/ash1 related protein 5 antibody
    • TRX5 antibody
    • TRX5 protein antibody
    • WHS antibody
    • Whsc1 antibody
    • Wolf Hirschhorn syndrome candidate 1 antibody
    • Wolf Hirschhorn syndrome candidate 1 protein antibody
    • Wolf-Hirschhorn syndrome candidate 1 protein antibody
    see all

Anti-WHSC1/NSD2 antibody [29D1] - ChIP Grade 图像

  • All lanes : Anti-WHSC1/NSD2 antibody [29D1] - ChIP Grade (ab75359) at 5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 152 kDa
    Observed band size : 80,180 kDa (why is the actual band size different from the predicted?)


    Exposure time : 20 minutes

    The full-length MMSET-II protein has an apparent molecular weight of approximately 180 kDa, whereas the 645 amino acid long MMSET-I exhibits a mobility of approximately 80 kDa.

     

    Abcam recommends blocking with 1% milk. 

  • IHC image of WHSC1/NSD2 staining in Human normal testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab75359, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • WHSC1/NSD2 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 10µg of Mouse monoclonal to WHSC1/NSD2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab75359.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 71kDa: WHSC1/NSD2.
  • All lanes : Anti-WHSC1/NSD2 antibody [29D1] - ChIP Grade (ab75359) at 1/5000 dilution

    Lane 1 : Whole cell lysate prepared from HeLa cells
    Lane 2 : Whole cell lysate prepared from HCT116 cells

    Lysates/proteins at 50 µg per lane.

    Secondary
    HRP conjugated goat anti-mouse polyclonal at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 152 kDa


    Exposure time : 3 minutes

    Image courtesy of an anonymous Abreview.

    Blocked with 5% milk for 1 hour at 23°C.

    See Abreview

  • ab75359 (1µg/ml) staining WHSC in human testis using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1/ in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

Anti-WHSC1/NSD2 antibody [29D1] - ChIP Grade (ab75359)参考文献

This product has been referenced in:
  • Hernando H  et al. EZH2 Inhibition Blocks Multiple Myeloma Cell Growth through Upregulation of Epithelial Tumor Suppressor Genes. Mol Cancer Ther 15:287-98 (2016). Read more (PubMed: 26590165) »
  • Saloura V  et al. WHSC1 promotes oncogenesis through regulation of NIMA-related kinase-7 in squamous cell carcinoma of the head and neck. Mol Cancer Res 13:293-304 (2015). WB, IP, IHC ; Human . Read more (PubMed: 25280969) »

See all 7 Publications for this product

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (BaF3 cell line NIH3T3 cell line HEK 293 cell line)
Specification BaF3 cell line NIH3T3 cell line HEK 293 cell line
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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提交于 Oct 01 2013

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (hct116, hela cells)
Loading amount 50 µg
Specification hct116, hela cells
Gel Running Conditions Reduced Denaturing (10% tris glycine)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

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Verified customer

提交于 Apr 30 2012

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"