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AB257565

Human NUDT1 (MTH1) knockout HEK-293T cell lysate

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NUDT1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 3.
3 Images
Western blot - Human NUDT1 (MTH1) knockout HEK-293T cell lysate (AB257565)
  • WB

Lab

Western blot - Human NUDT1 (MTH1) knockout HEK-293T cell lysate (AB257565)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : NUDT1 knockout HEK-293T cell lysate (20µg)

Lane 3 : HAP1 cell lysate (20µg)

Lane 4 : HeLa cell lysate (20µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab200832 observed at 18 kDa. Red - loading control ab8245 observed at 37 kDa.

ab200832 Anti-MTH1 antibody [EPR15934-50] was shown to specifically react with MTH1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266400 (knockout cell lysate ab257565) was used. Wild-type and MTH1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab200832 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 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-MTH1 antibody [EPR15934-50] (<a href='/en-us/products/primary-antibodies/mth1-antibody-epr15934-50-ab200832'>ab200832</a>) at 1/5000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

NUDT1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human NUDT1 (MTH1) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-nudt1-mth1-knockout-hek-293t-cell-line-ab266400'>ab266400</a>)

Lane 3:

HAP1 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Predicted band size: 23 kDa

Observed band size: 18 kDa

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Sanger Sequencing - Human NUDT1 (MTH1) knockout HEK-293T cell lysate (AB257565)
  • Sanger seq

Unknown

Sanger Sequencing - Human NUDT1 (MTH1) knockout HEK-293T cell lysate (AB257565)

Homozygous : 1 bp deletion in exon 3

Western blot - Human NUDT1 (MTH1) knockout HEK-293T cell lysate (AB257565)
  • WB

Lab

Western blot - Human NUDT1 (MTH1) knockout HEK-293T cell lysate (AB257565)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : NUDT1 knockout HEK-293T cell lysate (20µg)

Lane 3 : HAP1 cell lysate (20µg)

Lane 4 : HeLa cell lysate (20µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab197028 observed at 18 kDa. Red - loading control ab8245 observed at 37 kDa.

ab197028 Anti-MTH1 antibody [EPR15934] was shown to specifically react with MTH1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266400 (knockout cell lysate ab257565) was used. Wild-type and MTH1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab197028 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 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-MTH1 antibody [EPR15934] (<a href='/en-us/products/primary-antibodies/mth1-antibody-epr15934-ab197028'>ab197028</a>) at 1/2000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human NUDT1 (MTH1) knockout HEK-293T cell lysate (ab257565) at 20 µg

Lane 3:

HAP1 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Predicted band size: 23 kDa

Observed band size: 18 kDa

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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, Homozygous: 1 bp deletion in exon 3.

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Complementary Products

Primary Antibodies

AB200832

Anti-MTH1 antibody [EPR15934-50]

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTH1 antibody [EPR15934-50] (AB200832)

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  • 0
Proteins & Peptides

AB99390

Recombinant Human MTH1 protein

SDS-PAGE - Recombinant Human MTH1 protein (AB99390)

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Cell Lines & Lysates

AB266400

Human NUDT1 (MTH1) knockout HEK-293T cell line

Advanced Validation

Western blot - Human NUDT1 (MTH1) knockout HEK-293T cell line (AB266400)

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  • 0
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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 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
NUDT1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
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 target MTH1 also known as NUDT1 or karonudib is a protein with a mass of approximately 18.5 kDa. This protein belongs to the Nudix hydrolase family and is expressed in various tissues including liver brain and pancreas. Mechanically MTH1 plays a role in hydrolyzing oxidized nucleotides like 8-oxo-dGTP and 2-OH-dATP preventing their incorporation into DNA and maintaining genomic integrity.
Biological function summary

MTH1 prevents the incorporation of oxidized purine nucleotides into DNA. Without MTH1 cells risk mutations that can lead to genomic instability. The protein does not form part of a larger complex but acts independently. MTH1 functions are particularly relevant in cell types with high oxidative stress or rapidly dividing cells where its activity helps to protect against oxidative damage.

Pathways

MTH1 fits into DNA repair and antioxidant defense systems. It is notably involved in pathways managing oxidative stress damage. MTH1 works alongside other proteins like superoxide dismutase and catalase contributing to the detoxification of reactive oxygen species ultimately protecting the DNA repair machinery from oxidative damage.

MTH1 relates to cancer and neurodegenerative diseases. High MTH1 activity can be seen in cancer cells as these cells depend on MTH1 to prevent DNA damage from oxidative stress. In this context MTH1 pairs up with proteins like p53 which also has a role in maintaining genomic stability. In neurodegenerative diseases such as Parkinson's oxidative stress plays an important role implicating MTH1 since it mitigates oxidative nucleotide damage that might worsen neuronal damage.
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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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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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