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AB266758

Human SLC29A1 (ENT1) knockout HEK-293T cell line

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SLC29A1 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 4. 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 SLC29A1 (ENT1) knockout HEK-293T cell line (AB266758)
  • Sanger seq

Supplier Data

Sanger Sequencing - Human SLC29A1 (ENT1) knockout HEK-293T cell line (AB266758)

Homozygous : 1 bp insertion in exon 4

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 4

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Sanger seq": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.

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
SLC29A1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
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.

ENT1 also known as Equilibrative Nucleoside Transporter 1 or SLC29A1 is a membrane protein responsible for transporting nucleosides across cell membranes. This protein weighs approximately 55 kDa and is expressed in various tissues including the brain heart and liver. The transport activity of ENT1 plays a fundamental role in maintaining cellular nucleoside levels. The efficient transport mechanisms of ENT1 highlight its significance in regulating the availability of substrates for nucleic acid synthesis.
Biological function summary

This transporter facilitates the uptake and recycling of nucleosides which are essential building blocks for DNA and RNA synthesis. ENT1 mediates the transport of adenosine an important nucleoside involved in multiple cellular functions. It operates as a component of a broader transport system that includes similar proteins like ENT2. This action contributes to cellular processes by controlling nucleoside concentrations impacting cell proliferation and metabolic activities.

Pathways

ENT1 plays a role in the purine salvage pathway and adenosine signaling. It regulates extracellular adenosine levels influencing adenosine receptor-mediated pathways that impact many physiological functions. ENT1 coordinates with proteins such as adenosine kinases to manage the intracellular balance of adenosine therefore integrating into both metabolic and signaling routes. Its role in these pathways affects energy metabolism and supports cellular responses to stress.

ENT1 has been associated with conditions such as cardiovascular diseases and neurological disorders. Altered ENT1 activity can affect adenosine levels leading to variations in blood flow and neurotransmission. In cardiovascular diseases ENT1 modulates adenosine's cardioprotective effects. In neurological contexts dysfunction of ENT1 relates to disorders involving adenosine receptor signaling where its interaction with proteins like ADA (adenosine deaminase) can influence disease progression.

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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