Anti-nmt55 / p54nrb antibody (ab45359)
Key features and details
- Rabbit polyclonal to nmt55 / p54nrb
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-nmt55 / p54nrb antibody
See all nmt55 / p54nrb primary antibodies -
Description
Rabbit polyclonal to nmt55 / p54nrb -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Cow -
Immunogen
Synthetic peptide corresponding to Human nmt55/ p54nrb aa 1-100 (N terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab45358) -
Positive control
- ab45359 gave a positive result in the following whole cell lysates: Hela, A431, HEK293, HepG2, MCF7, U20S. ab45359 also gave a positive result in Hela Nuclear lysate.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentrationConcentration information loading...
Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab45359 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 1 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 54 kDa). IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. ICC/IF Use a concentration of 1 µg/ml. Target
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Function
DNA- and RNA binding protein, involved in several nuclear processes. Binds the conventional octamer sequence in double stranded DNA. Also binds single-stranded DNA and RNA at a site independent of the duplex site (By similarity). Involved in pre-mRNA splicing, probably as an heterodimer with SFPQ. Interacts with U5 snRNA, probably by binding to a purine-rich sequence located on the 3' side of U5 snRNA stem 1b. The SFPQ-NONO heteromer associated with MATR3 may play a role in nuclear retention of defective RNAs. The SFPQ-NONO heteromer may be involved in DNA unwinding by modulating the function of topoisomerase I/TOP1. The SFPQ-NONO heteromer may be involved in DNA nonhomologous end joining (NHEJ) required for double-strand break repair and V(D)J recombination and may stabilize paired DNA ends. In vitro, the complex strongly stimulates DNA end joining, binds directly to the DNA substrates and cooperates with the Ku70/G22P1-Ku80/XRCC5 (Ku) dimer to establish a functional preligation complex. Nono is involved in transcriptional regulation. The SFPQ-NONO-NR5A1 complex binds to the CYP17 promoter and regulates basal and cAMP-dependent transcriptional avtivity. NONO binds to an enhancer element in long terminal repeats of endogenous intracisternal A particles (IAPs) and activates transcription. -
Tissue specificity
Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. Also found in a number of breast tumor cell lines. -
Involvement in disease
Note=A chromosomal aberration involving NONO may be a cause of papillary renal cell carcinoma (PRCC). Translocation t(X;X)(p11.2;q13.1) with TFE3. -
Sequence similarities
Contains 2 RRM (RNA recognition motif) domains. -
Post-translational
modificationsThe N-terminus is blocked. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 615175 Cow
- Entrez Gene: 4841 Human
- Entrez Gene: 53610 Mouse
- Entrez Gene: 317259 Rat
- Omim: 300084 Human
- SwissProt: Q15233 Human
- SwissProt: Q99K48 Mouse
- SwissProt: Q5FVM4 Rat
see all -
Alternative names
- 52 kDa subunit antibody
- 54 kDa nuclear RNA and DNA binding protein antibody
- 54 kDa nuclear RNA- and DNA-binding protein antibody
see all
Images
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All lanes : Anti-nmt55 / p54nrb antibody (ab45359) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate
Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 7 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa. We are unsure as to the identity of these extra bands. -
ICC/IF image of ab45359 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab45359, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nmt55 / p54nrb antibody (ab45359)IHC image of nmt55/p54nrb staining in human breast carcinoma FFPE 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 ab45359, 1µg/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.
Protocols
Datasheets and documents
References (2)
ab45359 has been referenced in 2 publications.
- Torres M et al. Circadian RNA expression elicited by 3'-UTR IRAlu-paraspeckle associated elements. Elife 5:N/A (2016). IP, ICC/IF ; Rat . PubMed: 27441387
- Li Z et al. The long noncoding RNA THRIL regulates TNFa expression through its interaction with hnRNPL. Proc Natl Acad Sci U S A 111:1002-7 (2014). WB ; Human . PubMed: 24371310
Images
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All lanes : Anti-nmt55 / p54nrb antibody (ab45359) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate
Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 7 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa. We are unsure as to the identity of these extra bands.
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ICC/IF image of ab45359 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab45359, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nmt55 / p54nrb antibody (ab45359)IHC image of nmt55/p54nrb staining in human breast carcinoma FFPE 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 ab45359, 1µg/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.