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AB266740

Human NT5C knockout HEK-293T cell line

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NT5C KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 25 bp deletion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
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Sanger Sequencing - Human NT5C knockout HEK-293T cell line (AB266740)
  • Sanger seq

Unknown

Sanger Sequencing - Human NT5C knockout HEK-293T cell line (AB266740)

Homozygous : 25 bp deletion in exon 1

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: 25 bp deletion in exon 1

Product details

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?

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Properties and storage information

Gene name
NT5C
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.

NT5C also known as dNT-1 is a cytosolic enzyme with a molecular weight of approximately 41 kDa. It functions by dephosphorylating nucleoside monophosphates into nucleosides and its activity is important for nucleotide metabolism in cells. NT5C is expressed in various tissues including liver brain and skeletal muscle enabling these tissues to efficiently manage nucleotide balance.
Biological function summary

NT5C plays a role in maintaining cellular nucleotide pools impacting DNA and RNA synthesis. The enzyme is not part of a larger protein complex and primarily functions independently in nucleotide salvage pathways. Its activity influences the availability of nucleosides which are necessary for cellular replication and repair processes.

Pathways

NT5C contributes to purine and pyrimidine metabolism. It is involved in the nucleotide salvage pathways collaborating with other enzymes like adenosine deaminase and cytidine deaminase. These pathways play essential roles in recycling nucleotides and maintaining the nucleoside balance critical for cellular function.

NT5C has implications in cancer and neurological diseases. Abnormal activity of NT5C might contribute to nucleotide imbalances observed in certain cancers. Its role in dephosphorylation in the brain links it with neurodegenerative conditions potentially involving proteins like adenosine kinase. Understanding its function may offer insights into therapeutic strategies for these 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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