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

Rabbit Recombinant Monoclonal nmt55 / p54nrb antibody. Suitable for IHC-P, WB, ICC/IF and reacts with Mouse, Rat, Human samples. Cited in 7 publications.

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Images

Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (AB133574), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	WB	ICC/IF
Human	Tested	Tested	Tested
Mouse	Tested	Tested	Expected
Rat	Tested	Tested	Expected

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/250 - 1/500	Notes Use a HRP/AP polymerized secondary antibody. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Rat	Dilution info 1/250 - 1/500	Notes Use a HRP/AP polymerized secondary antibody. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Human	Dilution info 1/250 - 1/500	Notes Use a HRP/AP polymerized secondary antibody. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000 - 1/10000	Notes -
Species Rat	Dilution info 1/1000 - 1/10000	Notes -
Species Human	Dilution info 1/1000 - 1/10000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1500	Notes For unpurified use at 1/250 - 1/500.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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Target data

See full target information NONO

Function

DNA- and RNA binding protein, involved in several nuclear processes (PubMed:11525732, PubMed:12403470, PubMed:26571461). Binds the conventional octamer sequence in double-stranded DNA (PubMed:11525732, PubMed:12403470, PubMed:26571461). Also binds single-stranded DNA and RNA at a site independent of the duplex site (PubMed:11525732, PubMed:12403470, PubMed:26571461). Involved in pre-mRNA splicing, probably as a heterodimer with SFPQ (PubMed:11525732, PubMed:12403470, PubMed:26571461). Interacts with U5 snRNA, probably by binding to a purine-rich sequence located on the 3' side of U5 snRNA stem 1b (PubMed:12403470). Together with PSPC1, required for the formation of nuclear paraspeckles (PubMed:22416126). The SFPQ-NONO heteromer associated with MATR3 may play a role in nuclear retention of defective RNAs (PubMed:11525732). The SFPQ-NONO heteromer may be involved in DNA unwinding by modulating the function of topoisomerase I/TOP1 (PubMed:10858305). The SFPQ-NONO heteromer may be involved in DNA non-homologous end joining (NHEJ) required for double-strand break repair and V(D)J recombination and may stabilize paired DNA ends (PubMed:15590677). 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 (PubMed:15590677). NONO is involved in transcriptional regulation. The SFPQ-NONO-NR5A1 complex binds to the CYP17 promoter and regulates basal and cAMP-dependent transcriptional activity (PubMed:11897684). NONO binds to an enhancer element in long terminal repeats of endogenous intracisternal A particles (IAPs) and activates transcription (By similarity). Regulates the circadian clock by repressing the transcriptional activator activity of the CLOCK-BMAL1 heterodimer (By similarity). Important for the functional organization of GABAergic synapses (By similarity). Plays a specific and important role in the regulation of synaptic RNAs and GPHN/gephyrin scaffold structure, through the regulation of GABRA2 transcript (By similarity). Plays a key role during neuronal differentiation by recruiting TET1 to genomic loci and thereby regulating 5-hydroxymethylcytosine levels (By similarity). Plays a role in the regulation of DNA virus-mediated innate immune response by assembling into the HDP-RNP complex, a complex that serves as a platform for IRF3 phosphorylation and subsequent innate immune response activation through the cGAS-STING pathway (PubMed:28712728, PubMed:30270045). Promotes activation of the cGAS-STING pathway in response to HIV-2 infection: acts by interacting with HIV-2 Capsid protein p24, thereby promoting detection of viral DNA by CGAS, leading to CGAS-mediated inmmune activation (PubMed:30270045). In contrast, the weak interaction with HIV-1 Capsid protein p24 does not allow activation of the cGAS-STING pathway (PubMed:30270045).

Alternative names

NRB54, NONO, Non-POU domain-containing octamer-binding protein, NonO protein, 54 kDa nuclear RNA- and DNA-binding protein, 55 kDa nuclear protein, p54(nrb), p54nrb, NMT55

Recommended products

Rabbit Recombinant Monoclonal nmt55 / p54nrb antibody. Suitable for IHC-P, WB, ICC/IF and reacts with Mouse, Rat, Human samples. Cited in 7 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR5270
Purification technique: Affinity purification Protein A
Dissociation constant: 2.2 x 10^-12 M
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Stable for 12 months at -20°C

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The NMT55/p54nrb also known as the NonO protein is highly involved in various cellular processes. Mechanically it functions as a transcriptional coactivator and is involved in pre-mRNA splicing. This protein has a mass of approximately 54 kDa and is part of the Drosophila behaviour/human splicing (DBHS) protein family. It is ubiquitously expressed across different tissues with particular abundance in the nucleus showing its importance in gene regulation.

Biological function summary

P54nrb is a multifunctional protein critical in nuclear processes. It forms part of a complex involved in RNA processing and nuclear retention of edited RNA. This complex also includes the proteins PSF and PSPC1 contributing to its role in transcription regulation and RNA stability. p54nrb impacts gene expression through its ability to bind to DNA and RNA influencing numerous regulatory pathways within the cell.

Pathways

P54nrb has essential roles in gene expression and RNA maturation pathways. It closely interacts with the RNA polymerase II machinery affecting transcription elongation. The protein has an established connection with the steroid receptor RNA activator (SRA) to modulate transcriptional responses. It is related to other DBHS proteins such as PSF which work together for coordinate regulation of gene expression and splice site selection.

Associated diseases and disorders

P54nrb has been implicated in prostate cancer and amyotrophic lateral sclerosis (ALS). In prostate cancer altered expression or function of p54nrb can affect tumorigenesis sometimes through interactions with androgen receptor signaling pathways. Additionally in ALS mutations or defects in RNA processing involving p54nrb and FUS proteins lead to neurodegenerative processes. This relationship highlights its importance in maintaining normal cellular function and its impact when dysregulated.

Product promise

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13 product images

Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)
Lanes 1-3: Merged signal (red and green). Green - ab133574 observed at 63 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - 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 Human NONO (nmt55 / p54nrb) knockout HEK-293T cell line ab266244 (knockout cell lysate Human NONO (nmt55 / p54nrb) knockout HEK-293T 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 (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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.
All lanes: Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: NONO knockout HEK293T cell lysate at 20 µg
Lane 3: MOLT-4 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 63 kDa
Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)
Nmt55 / p54nrb Western blot staining using rabbit Anti-nmt55 / p54nrb antibody
Lanes 1-3: Merged signal (red and green). Green - ab133574 observed at 63 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - 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 Human NONO (nmt55 / p54nrb) knockout HEK-293T cell line ab266244 (knockout cell lysate Human NONO (nmt55 / p54nrb) knockout HEK-293T 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 (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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.
All lanes: Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: NONO knockout HEK293T cell lysate at 20 µg
Lane 3: MOLT-4 cell lysate at 20 µg
Secondary
Lanes 1 - 3: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773)
Lanes 1 - 3: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776)
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 63 kDa, 37 kDa
Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/10000 dilution
All lanes: PC-12 cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution
Predicted band size: 54 kDa
Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/10000 dilution
All lanes: RAW264.7 cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution
Predicted band size: 54 kDa
Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/10000 dilution
Lane 1: Molt-4 cell lysate at 20 µg
Lane 2: A673 cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution
Predicted band size: 54 kDa
Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)
All lanes: Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/1000 dilution
Lane 1: PC3 cell lysate at 10 µg
Lane 2: MOLT4 cell lysate at 10 µg
Lane 3: HeLa cell lysate at 10 µg
Lane 4: MCF7 cell lysate at 10 µg
Lane 5: A673 cell lysate at 10 µg
Lane 6: RAW 264.7 cell lysate at 10 µg
Lane 7: PC12 cell lysate at 10 µg
Lane 8: NIH/3T3 cell lysate at 10 µg
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. Goat Anti-Rabbit IgG H&L (HRP) 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. Goat Anti-Rabbit IgG H&L (HRP) 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. Goat Anti-Rabbit IgG H&L (HRP) 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. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).
OI-RD Scanning - Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574)
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.