Human NUDT5 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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NUDT5 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
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
- WB
Lab
Western blot - Human NUDT5 knockout HEK-293T cell line (AB266242)
Lanes 1-4 : Merged signal (red and green). Green - ab129163 observed at 30 kDa. Red - loading control ab8245 observed at 36 kDa.
ab129163 Anti-NUDT5 antibody [EPR7735] 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 (ab266242)
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
- Cell Culture
Lab
Cell Culture - Human NUDT5 knockout HEK-293T cell line (AB266242)
Representative images NUDT5 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 NUDT5 knockout HEK-293T cell line (AB266242)
Homozygous : Insertion of the selection cassette in exon2
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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.
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
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.
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
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NUDT5 antibody [EPR7734] (AB129172)](https://content.abcam.com/products/images/nudt5-antibody-epr7734-ab129172--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img126939.jpg)
Cell Lines & Lysates
AB266340
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Advanced Validation
- 0
- 0
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