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AB266650

Human GPX1 (Glutathione Peroxidase 1) knockout HEK-293T cell line

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GPX1 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.

View Alternative Names

AL033363, Cellular glutathione peroxidase, GPX1_HUMAN, GPXD, GSHPx-1, Glutathione peroxidase, Glutathione peroxidase 1, MGC14399, MGC88245

3 Images
Western blot - Human GPX1 (Glutathione Peroxidase 1) knockout HEK-293T cell line (AB266650)
  • WB

Lab

Western blot - Human GPX1 (Glutathione Peroxidase 1) knockout HEK-293T cell line (AB266650)

False colour image of Western blot : Anti-Glutathione Peroxidase 1 antibody [EPR3312] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab108427 was shown to bind specifically to Glutathione Peroxidase 1. A band was observed at 22 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in GPX1 knockout cell line ab266650 (knockout cell lysate ab256932). To generate this image wild-type and GPX1 knockout HEK-293T cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-Glutathione Peroxidase 1 antibody [EPR3312] (<a href='/en-us/products/primary-antibodies/glutathione-peroxidase-1-antibody-epr3312-ab108427'>ab108427</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

GPX1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human GPX1 (Glutathione Peroxidase 1) knockout HEK-293T cell line (ab266650)

Predicted band size: 22 kDa

Observed band size: 22 kDa

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Western blot - Human GPX1 (Glutathione Peroxidase 1) knockout HEK-293T cell line (AB266650)
  • WB

Lab

Western blot - Human GPX1 (Glutathione Peroxidase 1) knockout HEK-293T cell line (AB266650)

False colour image of Western blot : Anti-Glutathione Peroxidase 1 antibody [EPR3311] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab108429 was shown to bind specifically to Glutathione Peroxidase 1. A band was observed at 22 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in GPX1 knockout cell line ab266650 (knockout cell lysate ab256932). To generate this image wild-type and GPX1 knockout HEK-293T cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (<a href='/en-us/products/primary-antibodies/glutathione-peroxidase-1-antibody-epr3311-ab108429'>ab108429</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

GPX1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human GPX1 (Glutathione Peroxidase 1) knockout HEK-293T cell line (ab266650)

Predicted band size: 22 kDa

Observed band size: 22 kDa

false

Sanger Sequencing - Human GPX1 (Glutathione Peroxidase 1) knockout HEK-293T cell line (AB266650)
  • Sanger seq

Unknown

Sanger Sequencing - Human GPX1 (Glutathione Peroxidase 1) knockout HEK-293T cell line (AB266650)

Homozygous : 1 bp insertion 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: 1 bp insertion in exon 1

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Reactivity data

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

Recommended control: Human wild-type HEK293T 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.

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What's included?

{ "values": { "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab266650-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab266650 Human GPX1 (Glutathione Peroxidase 1) knockout HEK-293T cell line", "number":"AB266650-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
GPX1
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
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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.

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Supplementary information

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

Glutathione Peroxidase 1 (GPx1) also known as GSH peroxidase is a selenium-containing enzyme with a mass of approximately 22 kDa. It plays an important role in reducing hydrogen peroxide to water using glutathione as a substrate therefore protecting cells from oxidative damage. GPx1 is widely expressed in many tissues but is found in high concentrations in the liver and erythrocytes. Researchers use antibodies specific to GPx1 often obtained from specialized suppliers to study and quantify this enzyme in various biological samples.
Biological function summary

The enzyme guards cellular components by neutralizing free radicals. GPx1 is not part of a large complex but functions as a homotetramer. Its activity is essential for preserving the redox balance within the cell preventing cellular damage from reactive oxygen species (ROS). Enzyme-linked immunosorbent assays (ELISA) often measure GPx1 activity to assess oxidative stress levels in research and clinical settings.

Pathways

GPx1 is important in the glutathione metabolism and cellular antioxidant defense pathways. Within these pathways GPx1 works closely with proteins like glutathione reductase to maintain reduced glutathione levels. This interplay with other antioxidant proteins ensures cellular protection against oxidative damage. Studies focus on GPx1's role in these pathways as it underpins many critical cellular processes maintaining cell health.

Decreased GPx1 activity links to neurodegenerative diseases and cardiovascular disorders. In neurodegenerative diseases such as Alzheimer's the oxidative stress overwhelms the antioxidant defenses including GPx1. Similarly in cardiovascular disorders a diminished GPx1 activity correlates with increased oxidative stress contributing to disease progression. Researchers also investigate connections between GPx1 and other oxidative stress-related proteins like superoxide dismutase to fully understand GPx1's role in disease mechanisms.
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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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Target data

See full target information Glutathione peroxidase 1
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