Human NUDT1 (MTH1) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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NUDT1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 3.
View Alternative Names
2-hydroxy-dATP diphosphatase, 7 8 dihydro 8 oxoguanine triphosphatase, 8 oxo 7 8 dihydrodeoxyguanosine triphosphatase, 8 oxo 7 8 dihydroguanosine triphosphatase, 8-dihydro-8-oxoguanine triphosphatase, 8-oxo-dGTPase, 8ODP_HUMAN, MutT human homolog 1, NUDT 1, Nucleoside diphosphate linked moiety X type motif 1, Nucleoside diphosphate-linked moiety X motif 1, Nudix (nucleoside diphosphate linked moiety X) type motif 1, Nudix hydrolase 1, Nudix motif 1, Nudix type motif 1
- WB
Lab
Western blot - Human NUDT1 (MTH1) knockout HEK-293T cell line (AB266400)
Lanes 1- 4 : Merged signal (red and green). Green - ab197028 observed at 18 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab197028 was shown to 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 HEK-293T and NUDT1 knockout HEK-293T cell lysates 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) overnight at 4°C at a 1 in 2000 dilution and a 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:
NUDT1 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human NUDT1 (MTH1) knockout HEK-293T cell line (ab266400)
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
false
- WB
Lab
Western blot - Human NUDT1 (MTH1) knockout HEK-293T cell line (AB266400)
Lanes 1- 4 : Merged signal (red and green). Green - ab200832 observed at 18 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab200832 was shown to 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 HEK-293T and NUDT1 knockout HEK-293T cell lysates 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) overnight at 4°C at a 1 in 5000 dilution and a 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 (ab266400)
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
false
- Cell Culture
Lab
Cell Culture - Human NUDT1 (MTH1) knockout HEK-293T cell line (AB266400)
Representative images NUDT1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human NUDT1 (MTH1) knockout HEK-293T cell line (AB266400)
Homozygous : 1 bp deletion in exon 3
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
![Western blot - Anti-MTH1 antibody [EPR15934-50] (AB200832)](https://content.abcam.com/products/images/mth1-antibody-epr15934-50-ab200832--western-blot-img60971.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com