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AB257160

Human NONO (nmt55 / p54nrb) knockout HEK-293T cell lysate

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NONO KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon2 and 25 bp deletion in exon2.

View Alternative Names

52 kDa subunit, 54 kDa nuclear RNA- and DNA-binding protein, 55 kDa nuclear protein, DNA binding p52/p100 complex 52 kDa subunit, DNA-binding p52/p100 complex, NMT 55, NONO_HUMAN, NRB, NRB 54, Non POU domain containing octamer binding, Non Pou domain containing octamer (ATGCAAAT) binding protein, Non-POU domain-containing octamer-binding protein, NonO protein, Nuclear RNA binding protein 54kD, P54, PPP1R114, Protein phosphatase 1 regulatory subunit 114, p54(nrb), p54nrb

4 Images
Western blot - Human NONO (nmt55 / p54nrb) knockout HEK-293T cell lysate (AB257160)
  • WB

Unknown

Western blot - Human NONO (nmt55 / p54nrb) knockout HEK-293T cell lysate (AB257160)

Lane 1 : Wild-type HEK293T cell lysate (20 μg)

Lane 2 : NONO knockout HEK293T cell lysate (20 μg)

Lane 3 : MOLT-4 cell lysate (20 μg)

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.

All lanes:

Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (<a href='/en-us/products/primary-antibodies/nmt55-p54nrb-antibody-epr5270-ab133574'>ab133574</a>) 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

Predicted band size: 54 kDa

Observed band size: 63 kDa

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Sanger Sequencing - Human NONO (nmt55 / p54nrb) knockout HEK-293T cell lysate (AB257160)
  • Sanger seq

Unknown

Sanger Sequencing - Human NONO (nmt55 / p54nrb) knockout HEK-293T cell lysate (AB257160)

Allele-2 : 17 bp deletion in exon2

Sanger Sequencing - Human NONO (nmt55 / p54nrb) knockout HEK-293T cell lysate (AB257160)
  • Sanger seq

Unknown

Sanger Sequencing - Human NONO (nmt55 / p54nrb) knockout HEK-293T cell lysate (AB257160)

Allele-1 : 25 bp deletion in exon2

Western blot - Human NONO (nmt55 / p54nrb) knockout HEK-293T cell lysate (AB257160)
  • WB

Lab

Western blot - Human NONO (nmt55 / p54nrb) knockout HEK-293T cell lysate (AB257160)

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.

All lanes:

Western blot - Anti-nmt55 / p54nrb antibody [EPR5270] (<a href='/en-us/products/primary-antibodies/nmt55-p54nrb-antibody-epr5270-ab133574'>ab133574</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human NONO (nmt55 / p54nrb) knockout HEK-293T cell lysate (ab257160) 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 (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>)

Lanes 1 - 3:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>)

Predicted band size: 54 kDa

Observed band size: 63 kDa,37 kDa

false

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon2 and 25 bp deletion in exon2.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
NONO
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

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.

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.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Primary Antibodies

AB133574

Anti-nmt55 / p54nrb antibody [EPR5270]

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-nmt55 / p54nrb antibody [EPR5270] (AB133574)

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Product promise

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For full details, please see our Terms & Conditions

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