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GSS KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.

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Images

Western blot - Human GSS (Glutathione Synthetase) knockout HEK-293T cell line (AB266342), expandable thumbnail
  • Western blot - Human GSS (Glutathione Synthetase) knockout HEK-293T cell line (AB266342), expandable thumbnail
  • Sanger Sequencing - Human GSS (Glutathione Synthetase) knockout HEK-293T cell line (AB266342), expandable thumbnail

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

Knockout validation

Sanger Sequencing, Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2

Alternative names

GSH synthetase, GSH-S, GSHB_HUMAN, GSS, Glutathione synthase, Glutathione synthetase, HEL-S-64p, HEL-S-88n, MGC14098, OTTHUMP00000030711, epididymis secretory sperm binding protein Li 64p, epididymis secretory sperm binding protein Li 88n

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GSS KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.

Key facts

Cell type

HEK-293T

Form

Liquid

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2

Concentration
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Properties

Gene name

GSS

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Zygosity

Homozygous

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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.

Storage

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

Notes

Recommended control: Human wild-type HEK293T cell line (Human wild-type HEK-293T 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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Glutathione Synthetase (GSS) sometimes called GSS line is an enzyme that plays a central role in cellular function by catalyzing the ATP-dependent conversion of gamma-glutamylcysteine and glycine into glutathione. This enzyme has a molecular mass of about 70 kDa and exhibits high expression in the liver and red blood cells. It maintains the necessary cellular levels of glutathione a major antioxidant that protects cells from oxidative stress.

Biological function summary

GSS is responsible for the final step in glutathione biosynthesis. Acting independently GSS operates without forming a larger enzyme complex. Its enzymatic activity ensures adequate production of glutathione which regulates cellular redox homeostasis and detoxification processes. The availability of glutathione is critical for cellular respiration and immunity molecules essential for a well-functioning organism.

Pathways

The activity of GSS integrates into the gamma-glutamyl cycle and the detoxification pathway. It partners indirectly with enzymes like gamma-glutamylcysteine synthetase which precedes GSS in the glutathione biosynthesis pathway. This relationship positions GSS as an important element in sustaining normal cellular detoxification processes ensuring the neutralization of reactive oxygen species and maintaining the balance of free radicals.

Associated diseases and disorders

Defects in GSS activity correlate with glutathione synthetase deficiency an autosomal recessive disorder that results in increased oxidative damage and metabolic acidosis. Symptoms range from hemolytic anemia to neurological complications due to impairments in oxidative stress management. The disorder links to proteins involved in antioxidant processes highlighting the importance of GSS in maintaining proper cellular function and disease prevention.

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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