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AB266115

Human SEPHS1 (Selenophosphate synthetase 1) knockout HEK-293T cell line

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SEPHS1 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 2. 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 SEPHS1 (Selenophosphate synthetase 1) knockout HEK-293T cell line (AB266115)
  • Sanger seq

Unknown

Sanger Sequencing - Human SEPHS1 (Selenophosphate synthetase 1) knockout HEK-293T cell line (AB266115)

Homozygous : 1 bp insertion in exon 2

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 2

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?

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Properties and storage information

Gene name
SEPHS1
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.

Selenophosphate synthetase 1 also known as SEPHS1 or SPS1 catalyzes the synthesis of selenophosphate from selenide and ATP. It plays a critical role in the production of selenium-containing biomolecules. The mass of SEPHS1 approximates 46 kDa. SEPHS1 is expressed in various tissues including the liver and kidneys suggesting its involvement across multiple biological processes. Researchers often recognize SEPHS1 for its activity in selenium metabolism.
Biological function summary

The protein is involved in the synthesis of selenoproteins important for cellular antioxidant defense and redox homeostasis. Selenophosphate synthetase 1 functions as part of a multi-enzyme complex essential for the biosynthesis of selenocysteine the 21st amino acid which is then incorporated into selenoproteins. These selenoproteins include glutathione peroxidases and thioredoxin reductases which are important for maintaining cellular integrity.

Pathways

The protein participates actively in the selenocysteine biosynthesis and recycling pathway ensuring adequate supply of selenophosphate needed for selenoprotein production. Within this pathway SEPHS1 closely associates with several enzymes like selenocysteine synthase and selenocysteine tRNA (tRNASec) which facilitate the incorporation of selenocysteine into proteins. These interactions highlight the interconnected role of SEPHS1 in maintaining proper cellular functions through selenoproteins.

SEPHS1 is linked to deficiency-related conditions such as Keshan disease a type of heart disorder due to low selenium levels and certain thyroid dysfunctions linked to disrupted selenoprotein synthesis. Disruption of SEPHS1 function impairs selenoprotein production impacting proteins like glutathione peroxidase and thioredoxin reductase which are vital in preventing oxidative damage hence making SEPHS1 significant in understanding and potentially mitigating such disorders.

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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