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

Overview

  • Product name
    Anti-Nuclear Matrix Protein p84 antibody [5E10]
    See all Nuclear Matrix Protein p84 primary antibodies
  • Description
    Mouse monoclonal [5E10] to Nuclear Matrix Protein p84
  • Tested applications
    Suitable 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. WB: HEK-293T, A431, HeLa, HepG2, A-375 whole cell lysates; HeLa nuclear lysate. ICC/IF: HeLa cells. IP: HepG2 whole cell lysate. IHC-Fr: Human stomach tissue.

Properties

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.

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

Target

  • Function
    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.
  • Tissue specificity
    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.
  • Sequence similarities
    Contains 1 death domain.
  • Domain
    An 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 localization
    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
  • Database links
  • Form
    Nuclear (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

Images

  • All lanes : Anti-Nuclear Matrix Protein p84 antibody [5E10] (ab487) at 1/500 dilution

    Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 2 : A431 (human epidermoid carcinoma cell line) whole cell lysate
    Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 4 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 5 : A-375 (human malignant melanoma cell line) whole cell lysate

    Lysates/proteins at 30 µg per lane.

    Secondary
    HRP-conjugated anti-mouse IgG

    7.5% SDS-PAGE gel.

  • 4% paraformaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for Nuclear Matrix Protein p84 (green) using ab487 at 1/500 dilution in ICC/IF.

    Red: phalloidin, a cytoskeleton marker, at 1/200 dilution.

  • 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.

  • Nuclear Matrix Protein p84 was immunoprecipitated from HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate with 3 µg ab487. Western blot was performed from the immunoprecipitate using ab487. Anti-Rabbit IgG was used as a secondary reagent.

    Lane 1: HepG2 whole cell lysate 30 μg.

    Lane 2: Control IP in HepG2 whole cell lysate with 3 μg of pre-immune mouse IgG.

    Lane 3: ab487 IP in HepG2 whole cell lysate.

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

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) nuclear lysate

    Lysates/proteins at 30 µg per lane.

    Secondary
    HRP-conjugated anti-mouse IgG

    7.5% SDS-PAGE gel.

  • Acetone-fixed frozen section of human stomach tissue stained for Nuclear Matrix Protein p84 using ab487 at 2 µg/ml in immunohistochemical analysis.

    See Abreview

References

This product has been referenced in:
  • Zhang Y  et al. MicroRNA-410 promotes chondrogenic differentiation of human bone marrow mesenchymal stem cells through down-regulating Wnt3a. Am J Transl Res 9:136-145 (2017). WB ; Human . Read more (PubMed: 28123640) »
  • Alsafadi S  et al. Nuclear localization of the caspase-3-cleaved form of p73 in anoikis. Oncotarget 7:12331-43 (2016). WB . Read more (PubMed: 26575022) »

See all 46 Publications for this product

Customer reviews and Q&As

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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Abcam user community

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
Username

Abcam user community

Verified customer

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
Username

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
Username

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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Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

Submitted May 27 2010

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

Abcam user community

Verified customer

Submitted May 27 2010

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

Verified customer

Submitted May 12 2010

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

Verified customer

Submitted Mar 17 2010

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

Abcam user community

Verified customer

Submitted Sep 11 2009

1-10 of 13 Abreviews or Q&A

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