Anti-Nuclear Matrix Protein p84 antibody [5E10] (ab487)
Key features and details
- Mouse monoclonal [5E10] to Nuclear Matrix Protein p84
- Suitable for: WB, ICC/IF, IP, IHC-P, Flow Cyt
- Reacts with: Mouse, Rat, Human
- Isotype: IgG2b
Overview
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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 -
Host species
Mouse -
Tested applications
Suitable for: WB, ICC/IF, IP, IHC-P, Flow Cytmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Fusion protein containing amino acids 15-374 of human p84 expressed in E. coli.
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Positive control
- WB: HEK-293T, A431, HeLa, HepG2, A-375, NIH-3T3, PC-12 whole cell lysates; HeLa nuclear lysate, mouse cerebellum, brain tissue lysates. IHC-P: Human lung cencer and breast carcinoma tissue. ICC: HeLa cells. IP: HepG2 whole cell lysate.
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General notes
This product was changed from ascites to tissue culture supernatant on 2nd Feb 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.40
Constituent: 100% PBS -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
5E10 -
Myeloma
NS1 -
Isotype
IgG2b -
Light chain type
kappa -
Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
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 |
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WB | (8) |
Use a concentration of 0.3 - 2 µg/ml.
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ICC/IF | (2) |
Use a concentration of 0.5 - 2 µg/ml.
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IP | (1) |
Use at an assay dependent concentration.
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IHC-P | (2) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
Flow Cyt |
Use 1µg for 106 cells.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
Notes |
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WB
Use a concentration of 0.3 - 2 µg/ml. |
ICC/IF
Use a concentration of 0.5 - 2 µg/ml. |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt
Use 1µg for 106 cells. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
Target
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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
modificationsExpression 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
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Database links
- Entrez Gene: 9984 Human
- Entrez Gene: 225160 Mouse
- Entrez Gene: 291797 Rat
- Omim: 606930 Human
- SwissProt: Q96FV9 Human
- SwissProt: Q8R3N6 Mouse
- SwissProt: P59924 Rat
- Unigene: 712543 Human
see all -
Form
Nuclear (Isoform 1) and Cytoplasmic (Isoform 1 and 2). -
Alternative names
- hTREX84 antibody
- Death domain containing protein p84N5 antibody
- HPR 1 antibody
see all
Images
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Immunohistochemical analysis of human lung cancer tissue labeling Nuclear Matrix Protein p84 at the nucleus with ab487 at 1/100 dilution.
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All lanes : Anti-Nuclear Matrix Protein p84 antibody [5E10] (ab487) at 1/1000 dilution
Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate
Lane 3 : Mouse brain tissue lysate
Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : anti-mouse IgG HRP-conjugated antibody -
Immunocytochemical analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Nuclear Matrix Protein p84 at the nucleus with ab487 at 1/500 dilution. Red: phalloidin, a cytoskeleton marker, diluted at 1:100.
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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.
This image was generated using the ascites version of the product.
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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.
This image was generated using the ascites version of the product.
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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
All lanes : HRP-conjugated anti-mouse IgG7.5% SDS-PAGE gel.
This image was generated using the ascites version of the product.
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Immunohistochemical analysis of human breast carcinoma tissue labeling Nuclear Matrix Protein p84 at the nucleus with ab487 at 1/200 dilution.
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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.
This image was generated using the ascites version of the product.
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All lanes : Anti-Nuclear Matrix Protein p84 antibody [5E10] (ab487) at 1/1000 dilution
Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2 : HEK-293T whole cell lysate at 10 µg
Lane 3 : HEK-293T whole cell lysate at 5 µg
Secondary
All lanes : anti-mouse IgG HRP-conjugated antibody -
ICC/IF image of ab487 stained HepG2 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.
This image was generated using the ascites version of the product.
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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
All lanes : HRP-conjugated anti-mouse IgG7.5% SDS-PAGE gel.
This image was generated using the ascites version of the product.
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Anti-Nuclear Matrix Protein p84 antibody [5E10] (ab487) at 1/1000 dilution + Mouse cerebellum tissue lysates at 50 µg
Secondary
anti-mouse IgG HRP-conjugated antibody
Protocols
Datasheets and documents
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Datasheet download
References (90)
ab487 has been referenced in 90 publications.
- Polenkowski M et al. THOC5 complexes with DDX5, DDX17, and CDK12 to regulate R loop structures and transcription elongation rate. iScience 26:105784 (2023). PubMed: 36590164
- Sun YJ et al. HABP1 promotes proliferation and invasion of lung adenocarcinoma cells through NFκB pathway. Neoplasma 69:155-164 (2022). PubMed: 34881631
- El-Hindi K et al. T-Cell-Specific CerS4 Depletion Prolonged Inflammation and Enhanced Tumor Burden in the AOM/DSS-Induced CAC Model. Int J Mol Sci 23:N/A (2022). PubMed: 35163788
- Shin HJ et al. Lipocalin-2 Deficiency Reduces Hepatic and Hippocampal Triggering Receptor Expressed on Myeloid Cells-2 Expressions in High-Fat Diet/Streptozotocin-Induced Diabetic Mice. Brain Sci 12:N/A (2022). PubMed: 35884685
- de Mingo Pulido Á et al. The inhibitory receptor TIM-3 limits activation of the cGAS-STING pathway in intra-tumoral dendritic cells by suppressing extracellular DNA uptake. Immunity 54:1154-1167.e7 (2021). PubMed: 33979578