Anti-Nuclear Matrix Protein p84抗体[5E10] (ab487)

概述

  • 产品名称Anti-Nuclear Matrix Protein p84抗体[5E10]
    参阅全部 Nuclear Matrix Protein p84 一抗
  • 描述
    小鼠单克隆抗体[5E10] to核Matrix Protein p84
  • 经测试应用适用于: WB, IP, ICC/IF, ICC, IHC-P, IHC-Fr, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    Fusion protein containing amino acids 15-374 of human p84 expressed in E. coli.

  • 阳性对照
    • Molt 4, HeLa, Raji cells.

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • 存储溶液Preservative: None
    Constituents: 10mM PBS, pH 7.4
  • Concentration information loading...
  • 纯度Protein G purified
  • 纯化说明Purified from hybridoma cell culture supernatant by protein G chromatography to at least 95% homogeneity as determined by SDS-PAGE.
  • 克隆单克隆
  • 克隆编号5E10
  • 骨髓瘤NS1
  • 同种型IgG2b
  • 轻链类型kappa
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab487 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
WB Use a concentration of 0.5 - 2 µg/ml.
IP Use a concentration of 0.5 - 2 µg/ml.
ICC/IF Use a concentration of 0.5 - 2 µg/ml.
ICC Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

靶标

  • 功能Component of the THO subcomplex of the TREX complex. The TREX complex specifically associates with spliced mRNA and not with unspliced pre-mRNA. It is recruited to spliced mRNAs by a transcription-independent mechanism. Binds to mRNA upstream of the exon-junction complex (EJC) and is recruited in a splicing- and cap-dependent manner to a region near the 5' end of the mRNA where it functions in mRNA export. The recruitment occurs via an interaction between THOC4 and the cap-binding protein NCBP1. DDX39B functions as a bridge between THOC4 and the THO complex. The TREX complex is essential for the export of Kaposi's sarcoma-associated herpesvirus (KSHV) intronless mRNAs and infectious virus production. The recruitment of the TREX complex to the intronless viral mRNA occurs via an interaction between KSHV ORF57 protein and THOC4.
    Regulates transcriptional elongation of a subset of genes. Participates in an apoptotic pathway which is characterized by activation of caspase-6, increases in the expression of BAK1 and BCL2L1 and activation of NF-kappa-B. This pathway does not require p53/TP53, nor does the presence of p53/TP53 affect the efficiency of cell killing. Activates a G2/M cell cycle checkpoint prior to the onset of apoptosis. Apoptosis is inhibited by association with RB1.
  • 组织特异性Ubiquitous. Expressed in various cancer cell lines. Expressed at very low levels in normal breast epithelial cells and highly expressed in breast tumors. Expression is strongly associated with an aggressive phenotype of breast tumors and expression correlates with tumor size and the metastatic state of the tumor progression.
  • 序列相似性Contains 1 death domain.
  • 结构域An intact death domain is needed for apoptosis.
  • 翻译后修饰Expression is altered specifically during apoptosis and is accompanied by the appearance of novel forms with smaller apparent molecular mass.
  • 细胞定位Cytoplasm and Nucleus speckle. Nucleus > nucleoplasm. Nucleus matrix. Cytoplasm. Can shuttle between the nucleus and cytoplasm. Nuclear localization is required for induction of apoptotic cell death. Translocates to the cytoplasm during the early phase of apoptosis execution.
  • Information by UniProt
  • 数据库链接
  • 形式Nuclear (Isoform 1) and Cytoplasmic (Isoform 1 and 2).
  • 别名
    • hTREX84 antibody
    • Death domain containing protein p84N5 antibody
    • HPR 1 antibody
    • HPR1 antibody
    • hTREX84 antibody
    • Nuclear matrix protein p84 antibody
    • P84 antibody
    • p84N5 antibody
    • Tho 1 antibody
    • THO complex 1 antibody
    • THO complex subunit 1 antibody
    • Tho1 antibody
    • THOC 1 antibody
    • Thoc1 antibody
    • THOC1_HUMAN antibody
    see all

Anti-Nuclear Matrix Protein p84 antibody [5E10] 图像

  • All lanes : Anti-Nuclear Matrix Protein p84 antibody [5E10] (ab487) at 1/1000 dilution

    Lane 1 : 0.1% DMSO treated Jurkat cell (whole cell lysate)
    Lane 2 : 0.5% DMSO treated Jurkat cell (whole cell lysate)
    Lane 3 : 1% DMSO treated Jurkat cell (whole cell lysate)
    Lane 4 : 0.1% SDS treated Jurkat cell (whole cell lysate)
    Lane 5 : 0.5% SDS treated Jurkat cell (whole cell lysate)
    Lane 6 : 1% SDS treated Jurkat cell (whole cell lysate)

    Secondary
    HRP conjugated goat anti-mouse.
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 84 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    This image is courtesy of an anonymous Abreview

    SDS and DMSO were constituents of the lysis buffer. 0.1% SDS did not break down the nucleus entirely , however the higher concentrations did and p84 was detected in the lysate.

    See Abreview

  • ab487 staining Nuclear Matrix Protein p84 in Human stomach adenocarcinoma cell line (AGS) by Immunocytochemistry/ Immunofluorescence. The cells were formaldehyde fixed, permeabilised in 0.025% Triton X-100, TBS and then blocked using 5% serum for 1 hour at 23°C. Samples were then incubated with primary antibody at 2µg/ml for 1 hour at 23°C. The secondary antibody used was a goat anti-mouse IgG conjugated to Alexa Fluor® 350 (blue) used undiluted.
    p84 shows nuclear localization.

    See Abreview

  • ICC/IF image of ab487 stained Hepp 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 (ab487, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.

  • Overlay histogram showing HeLa cells stained with ab487 (red line). The cells were fixed with 100% methanol (5 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 (ab487, 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 IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

Anti-Nuclear Matrix Protein p84 antibody [5E10] (ab487)参考文献

This product has been referenced in:
  • Salzman DW  et al. miR-34 activity is modulated through 5'-end phosphorylation in response to DNA damage. Nat Commun 7:10954 (2016). WB ; Human . Read more (PubMed: 26996824) »
  • Meier D  et al. Twist1 Is a TNF-Inducible Inhibitor of Clock Mediated Activation of Period Genes. PLoS One 10:e0137229 (2015). WB . Read more (PubMed: 26361389) »

See all 31 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - nuclear (cervix)
Loading amount 50 µg
Specification cervix
Gel Running Conditions Reduced Denaturing (10)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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提交于 Feb 09 2012

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Application Western blot
Sample Mouse Cell lysate - whole cell (NSC34 cells)
Loading amount 10 µg
Specification NSC34 cells
Gel Running Conditions Reduced Denaturing (4-12% Nupage gel)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Jul 08 2011

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Application Western blot
Sample Human Cell lysate - whole cell (HeLa)
Loading amount 35 µg
Specification HeLa
Gel Running Conditions Reduced Denaturing (7% Tris-Acetat)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Dr. Lukas Rambousek

Verified customer

提交于 Dec 22 2010

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Application Western blot
Sample Human Cell lysate - nuclear (Fibroblasts)
Loading amount 40 µg
Specification Fibroblasts
Gel Running Conditions Reduced Denaturing (4-12% BisTris)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Dr. Ioannis Gavvovidis

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提交于 Dec 03 2010

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Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (Stomach)
Specification Stomach
Fixative Acetone
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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提交于 May 27 2010

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Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Stomach)
Specification Stomach
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10mM citrate pH6.0
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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提交于 May 27 2010

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Application Immunocytochemistry
Sample Human Cultured Cells (AGS Gastric Carcinoma cells)
Specification AGS Gastric Carcinoma cells
Fixative Formaldehyde
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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提交于 May 27 2010

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (AGS)
Specification AGS
Fixative Formaldehyde
Permeabilization Yes - 0.025% Triton-X in TBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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提交于 May 12 2010

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Application Immunoprecipitation
Sample Human Cell lysate - whole cell (AGS)
Total protein in input 200 µg
Specification AGS
Immuno-precipitation step Protein A/G
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提交于 Mar 17 2010

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Application Western blot
Sample Human Cell lysate - nuclear (Hela)
Loading amount 60 µg
Specification Hela
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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提交于 Sep 11 2009

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