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

Overview

  • Product nameAnti-Nuclear Matrix Protein p84 antibody [5E10]
    See all Nuclear Matrix Protein p84 primary antibodies
  • Description
    Mouse monoclonal [5E10] to Nuclear Matrix Protein p84
  • Tested applicationsSuitable for: WB, IP, ICC/IF, ICC, IHC-P, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

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

  • Positive control
    • Molt 4, HeLa, Raji cells.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: None
    Constituents: 10mM PBS, pH 7.4
  • Concentration information loading...
  • PurityProtein G purified
  • Purification notesPurified from hybridoma cell culture supernatant by protein G chromatography to at least 95% homogeneity as determined by SDS-PAGE.
  • ClonalityMonoclonal
  • Clone number5E10
  • MyelomaNS1
  • IsotypeIgG2b
  • Light chain typekappa
  • Research areas

Applications

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.

Application Abreviews Notes
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.

Target

  • FunctionComponent 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.
  • Tissue specificityUbiquitous. 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.
  • Sequence similaritiesContains 1 death domain.
  • DomainAn intact death domain is needed for apoptosis.
  • Post-translational
    modifications
    Expression is altered specifically during apoptosis and is accompanied by the appearance of novel forms with smaller apparent molecular mass.
  • Cellular localizationCytoplasm 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
  • Database links
  • FormNuclear (Isoform 1) and Cytoplasmic (Isoform 1 and 2).
  • Alternative names
    • 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] images

  • 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

    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.

References for Anti-Nuclear Matrix Protein p84 antibody [5E10] (ab487)

This product has been referenced in:
  • Stubbs SH & Conrad NK Depletion of REF/Aly alters gene expression and reduces RNA polymerase II occupancy. Nucleic Acids Res 43:504-19 (2015). Read more (PubMed: 25477387) »
  • Seiki T  et al. HPV-16 impairs the subcellular distribution and levels of expression of protein phosphatase 1? in cervical malignancy. BMC Cancer 15:230 (2015). Read more (PubMed: 25886518) »

See all 29 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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Verified customer

Submitted Feb 09 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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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Submitted Jul 08 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Submitted Dec 22 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Verified customer

Submitted Dec 03 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
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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Submitted May 27 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
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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Submitted 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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Submitted 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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Submitted 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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Submitted 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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Submitted Sep 11 2009

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