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AB267326

Human NADK knockout HEK-293T cell line

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NADK KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.

View Alternative Names

FLJ13052, FLJ37724, NAD kinase, NADK_HUMAN, PPNK, Poly(P)/ATP NAD kinase, RP1-283E3.6, dJ283E3.1

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Sanger Sequencing - Human NADK knockout HEK-293T cell line (AB267326)
  • Sanger seq

Unknown

Sanger Sequencing - Human NADK knockout HEK-293T cell line (AB267326)

Homozygous : 1 bp insertion in exon1

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1

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, 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?

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Properties and storage information

Gene name
NADK
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

NAD kinase also known as NADK is a critical enzyme responsible for phosphorylating nicotinamide adenine dinucleotide (NAD+) to form NADP+. NADK has an approximate molecular weight of 50 kDa. Its expression is widespread but it is especially abundant in pancreatic and liver tissues. This enzyme converts NAD+ to NADP+ which is important for maintaining the balance of NADP+/NADPH within cells impacting several biochemical processes.
Biological function summary

The conversion role played by NADK is essential for maintaining cellular redox status and for biosynthetic reactions that require NADPH such as fatty acid synthesis and detoxifying reactive oxygen species. NADK functions independently without being part of larger protein complexes. It facilitates key cellular processes by ensuring adequate levels of NADPH a necessary cofactor in various anabolic reactions and cellular defense mechanisms.

Pathways

NADK prominently features in the pentose phosphate pathway which is important for the production of NADPH. It interacts with enzymes like glucose-6-phosphate dehydrogenase (G6PD) which is also involved in producing NADPH through this pathway. Additionally NADK plays a role in the citric acid cycle where its activity supports energy metabolism by maintaining cellular NADP+/NADPH ratios.

Altered NADK activity links to conditions such as cancer and diabetes. Its role in regulating redox state and metabolic pathways means that dysregulation can lead to oxidative stress contributing to the pathogenesis of several types of cancer. Disrupted glucose metabolism in diabetes associates G6PD and NADK since both are pivotal in maintaining NADPH levels highlighting the enzyme's connection with metabolic diseases.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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