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AB257460

Human GSS (Glutathione Synthetase) knockout HEK-293T cell lysate

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GSS KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
3 Images
Western blot - Human GSS (Glutathione Synthetase) knockout HEK-293T cell lysate (AB257460)
  • WB

Lab

Western blot - Human GSS (Glutathione Synthetase) knockout HEK-293T cell lysate (AB257460)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : GSS knockout HEK-293T cell lysate (20µg)

Lane 3 : HeLa cell lysate (20µg)

Lane 4 : Daudi cell lysate (20µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab133592 observed at 50 kDa. Red - loading control ab8245 observed at 37 kDa.

ab133592 Anti-Glutathione Synthetase antibody [EPR6563] was shown to specifically react with GSS in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266342 (knockout cell lysate ab257460) was used. Wild-type and GSS knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133592 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Glutathione Synthetase antibody [EPR6563] (<a href='/en-us/products/primary-antibodies/glutathione-synthetase-antibody-epr6563-ab133592'>ab133592</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

GSS knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human GSS (Glutathione Synthetase) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-gss-glutathione-synthetase-knockout-hek-293t-cell-line-ab266342'>ab266342</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 52 kDa

Observed band size: 50 kDa

false

Western blot - Human GSS (Glutathione Synthetase) knockout HEK-293T cell lysate (AB257460)
  • WB

Lab

Western blot - Human GSS (Glutathione Synthetase) knockout HEK-293T cell lysate (AB257460)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : GSS knockout HEK-293T cell lysate (20µg)

Lane 3 : HeLa cell lysate (20µg)

Lane 4 : Daudi cell lysate (20µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab124811 observed at 52 kDa. Red - loading control ab8245 observed at 37 kDa.

ab124811 Anti-Glutathione Synthetase antibody [EPR6562] was shown to specifically react with GSS in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266342 (knockout cell lysate ab257460) was used. Wild-type and GSS knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab124811 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Glutathione Synthetase antibody [EPR6562] (<a href='/en-us/products/primary-antibodies/glutathione-synthetase-antibody-epr6562-ab124811'>ab124811</a>) at 1/10000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

GSS knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human GSS (Glutathione Synthetase) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-gss-glutathione-synthetase-knockout-hek-293t-cell-line-ab266342'>ab266342</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 52 kDa

Observed band size: 52 kDa

false

Sanger Sequencing - Human GSS (Glutathione Synthetase) knockout HEK-293T cell lysate (AB257460)
  • Sanger seq

Unknown

Sanger Sequencing - Human GSS (Glutathione Synthetase) knockout HEK-293T cell lysate (AB257460)

Homozygous : 1 bp insertion in exon 2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

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

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Complementary Products

Primary Antibodies

AB133592

Anti-Glutathione Synthetase antibody [EPR6563]

RabMAb|Recombinant|KO Validated

Flow Cytometry (Intracellular) - Anti-Glutathione Synthetase antibody [EPR6563] (AB133592)

  • 0
  • 0
Cell Lines & Lysates

AB266342

Human GSS (Glutathione Synthetase) knockout HEK-293T cell line

Advanced Validation

Western blot - Human GSS (Glutathione Synthetase) knockout HEK-293T cell line (AB266342)

  • 0
  • 0
Primary Antibodies

AB110296

Anti-ACADM/MCAD antibody [3B7BH7]

Immunocytochemistry/ Immunofluorescence - Anti-ACADM/MCAD antibody [3B7BH7] (AB110296)

  • 4
  • 1
Assay Kits

AB283966

Human Glutathione Synthetase ELISA Kit

Sandwich ELISA - Human Glutathione Synthetase ELISA Kit (AB283966)

  • 0
  • 0
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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
GSS
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

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.

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.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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