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AB258087

Human NUDT5 knockout HEK-293T cell lysate

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NUDT5 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Insertion of the selection cassette in exon2.

View Alternative Names

ADP-sugar pyrophosphatase, NUDT5_HUMAN, Nucleoside diphosphate linked moiety X type motif 5, Nucleoside diphosphate-linked moiety X motif 5, Nudix (nucleoside diphosphate linked moiety X) type motif 5, Nudix motif 5, Nudix type motif 5, YSA1, YSA1H, hYSAH 1

3 Images
Western blot - Human NUDT5 knockout HEK-293T cell lysate (AB258087)
  • WB

Lab

Western blot - Human NUDT5 knockout HEK-293T cell lysate (AB258087)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : NUDT5 knockout HEK293T cell lysate (20 ug)
Lane 3 : HepG2 cell lysate (20 ug)
Lane 4 : U-937 cell lysate (20 ug)

ab129172 was shown to specifically react with NUDT5 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266242 (knockout cell lysate ab258087) was used. Wild-type and NUDT5 knockout samples were subjected to SDS-PAGE. ab129172 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NUDT5 antibody [EPR7734] (<a href='/en-us/products/primary-antibodies/nudt5-antibody-epr7734-ab129172'>ab129172</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

NUDT5 knockout HEK293T cell lysate at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

U-937 cell lysate at 20 µg

Secondary

All lanes:

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>) at 1/10000 dilution

Predicted band size: 24 kDa

Observed band size: 30 kDa

false

Western blot - Human NUDT5 knockout HEK-293T cell lysate (AB258087)
  • WB

Lab

Western blot - Human NUDT5 knockout HEK-293T cell lysate (AB258087)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : NUDT5 knockout HEK293T cell lysate (20 ug)
Lane 3 : HepG2 cell lysate (20 ug)
Lane 4 : HeLa cell lysate (20 ug)

ab129163 was shown to specifically react with NUDT5 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266242 (knockout cell lysate ab258087) was used. Wild-type and NUDT5 knockout samples were subjected to SDS-PAGE. ab129163 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NUDT5 antibody [EPR7735] (<a href='/en-us/products/primary-antibodies/nudt5-antibody-epr7735-ab129163'>ab129163</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

NUDT5 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human NUDT5 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-nudt5-knockout-hek-293t-cell-line-ab266242'>ab266242</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

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>) at 1/10000 dilution

Predicted band size: 24 kDa

Observed band size: 30 kDa

false

Sanger Sequencing - Human NUDT5 knockout HEK-293T cell lysate (AB258087)
  • Sanger seq

Unknown

Sanger Sequencing - Human NUDT5 knockout HEK-293T cell lysate (AB258087)

Homozygous : Insertion of the selection cassette in exon2

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, Insertion of the selection cassette in exon2.

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Reactivity data

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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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
NUDT5
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 Nudix hydrolase 5 (NUDT5) enzyme also known as ADP-sugar pyrophosphatase performs important mechanical roles by hydrolyzing ADP-ribose to ribose-5-phosphate and AMP. It holds a molecular mass of approximately 39 kDa. NUDT5 shows high expression levels in tissues like liver and ovary. Its activity involves detoxifying nucleoside diphosphate derivatives which prevents their accumulation and potential interference with normal cellular processes.
Biological function summary

NUDT5 influences intracellular signaling pathways and DNA repair processes. It operates as part of multi-enzyme complexes that manage nucleotide metabolism and signaling. By maintaining a balance of ADP-ribose inside cells NUDT5 helps regulate cellular stress responses and genomic stability. Its role in transforming ADP-ribose supports maintaining adequate cellular energy and nucleotide pools for various biological functions.

Pathways

Research identifies NUDT5 as being involved in the ADP-ribose metabolism pathway and DNA repair mechanisms. In the ADP-ribose pathway NUDT5 assists in the regulation of ADP-ribose levels working alongside other proteins like PARP1 which adds ADP-ribose units to proteins during DNA repair. NUDT5's actions in these pathways ensure the proper functioning of cellular stress responses and the prevention of genome instability.

NUDT5 has connections to tumorigenesis and neurodegenerative disorders. Elevated activity of NUDT5 is sometimes observed in certain cancers suggesting a potential role in cancer cell viability. In neurodegenerative diseases such as Parkinson's dysfunctional ADP-ribose metabolism implicates NUDT5. Changes in NUDT5 activity can interact with proteins like PARP1 altering cellular processes that contribute to disease pathogenesis.
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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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