Recombinant Anti-nmt55 / p54nrb antibody [EPR5270] KO Tested (ab133574)

Product name

Anti-nmt55 / p54nrb antibody [EPR5270]
See all nmt55 / p54nrb primary antibodies
Description

Rabbit monoclonal [EPR5270] to nmt55 / p54nrb
Host species

Rabbit
Tested applications

Suitable for: WB, ICC/IF, IHC-Pmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide within Human nmt55/ p54nrb (C terminal). The exact sequence is proprietary.
Database link: Q15233
Positive control
- WB: HEK293T, PC3, MOLT4, HeLa, MCF7, A673, RAW 264.7, PC12 and NIH/3T3 cell lysates. IHC-P: Human kidney and bladder carcinoma tissues.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Dissociation constant (K_D)

K_D = 2.20 x 10 ^-12 M
Learn more about K_D
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR5270
Isotype

IgG
Research areas

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133574 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		1/1000 - 1/10000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
ICC/IF		1/1500. For unpurified use at 1/250 - 1/500.
IHC-P		1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. Use a HRP/AP polymerized secondary antibody.

Notes
WB 1/1000 - 1/10000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
ICC/IF 1/1500. For unpurified use at 1/250 - 1/500.
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. Use a HRP/AP polymerized secondary antibody.

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
modifications

The N-terminus is blocked.
Cellular localization

Nucleus.
Target information above from: UniProt accession Q15233 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 4841 Human
- Entrez Gene: 53610 Mouse
- Entrez Gene: 317259 Rat
- Omim: 300084 Human
- SwissProt: Q15233 Human
- SwissProt: Q99K48 Mouse
- SwissProt: Q5FVM4 Rat
- Unigene: 533282 Human
- Unigene: 700344 Human
- Unigene: 280069 Mouse
- Unigene: 8381 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
- 55 kDa nuclear protein antibody
- DNA binding p52/p100 complex 52 kDa subunit antibody
- DNA-binding p52/p100 complex antibody
- NMT 55 antibody
- NMT55 antibody
- Non Pou domain containing octamer (ATGCAAAT) binding protein antibody
- Non POU domain containing octamer binding antibody
- Non POU domain containing octamer binding protein antibody
- Non-POU domain-containing octamer-binding protein antibody
- Nono antibody
- NonO protein antibody
- NONO_HUMAN antibody
- NRB 54 antibody
- NRB antibody
- NRB54 antibody
- Nuclear RNA binding protein 54kD antibody
- P54 antibody
- p54(nrb) antibody
- p54nrb antibody
- PPP1R114 antibody
- Protein phosphatase 1 regulatory subunit 114 antibody
see all

Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)

All lanes : Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : NONO knockout HEK293T cell lysate
Lane 3 : MOLT-4 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 54 kDa
Observed band size: 63 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab133574 observed at 63 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab133574 Anti-nmt55 / p54nrb antibody [EPR5270] was shown to specifically react with nmt55 / p54nrb in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266244 (knockout cell lysate ab257160) was used. Wild-type and nmt55 / p54nrb knockout samples were subjected to SDS-PAGE. ab133574 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)

Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/10000 dilution (purified) + PC-12 cell lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 54 kDa

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)

Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/10000 dilution (purified) + RAW264.7 cell lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 54 kDa

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)

All lanes : Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/10000 dilution (purified)

Lane 1 : Molt-4 cell lysate
Lane 2 : A673 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 54 kDa

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)

All lanes : Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/1000 dilution (unpurified)

Lane 1 : PC3 cell lysate
Lane 2 : MOLT4 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : MCF7 cell lysate
Lane 5 : A673 cell lysate
Lane 6 : RAW 264.7 cell lysate
Lane 7 : PC12 cell lysate
Lane 8 : NIH/3T3 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 54 kDa
Observed band size: 54 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling nmt55/p54nrb with purified ab133574 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue labelling nmt55/p54nrb with purified ab133574 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate tissue labelling nmt55/p54nrb with purified ab133574 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human bladder carcinoma tissue labelling nmt55 / p54nrb using unpurified ab133574 at a 1/250 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human kidney tissue labelling nmt55 /p54nrb using unpurified ab133574 at a 1/250 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)

Immunocytochemistry/Immunofluorescence analysis of Molt-4 (human acute lymphoblastic leukemia) cells labelling nmt55/p54nrb with purified ab133574 at 1/1500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).
OI-RD Scanning - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)

Equilibrium disassociation constant (K_D)
Learn more about K_D

Click here to learn more about K_D
Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)

Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab133574? Please let us know so that we can cite the reference in this datasheet.

ab133574 has been referenced in 3 publications.

Shen W et al. Acute hepatotoxicity of 2' fluoro-modified 5-10-5 gapmer phosphorothioate oligonucleotides in mice correlates with intracellular protein binding and the loss of DBHS proteins. Nucleic Acids Res 46:2204-2217 (2018). PubMed: 29390093
Hupalowska A et al. CARM1 and Paraspeckles Regulate Pre-implantation Mouse Embryo Development. Cell 175:1902-1916.e13 (2018). PubMed: 30550788
Shadle SC et al. DUX4-induced dsRNA and MYC mRNA stabilization activate apoptotic pathways in human cell models of facioscapulohumeral dystrophy. PLoS Genet 13:e1006658 (2017). ICC/IF ; Human . PubMed: 28273136

Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Dissociation constant (KD)

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Isotype control

KO cell lines

KO cell lysates

Positive Controls

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

References (3)

Customer reviews and Q&As

Dissociation constant (K_D)

Post-translational
modifications