AB263935

Human GPX4 knockout HeLa cell lysate

Name: Human GPX4 knockout HeLa cell lysate (ab263935) | Abcam
Brand: Abcam Limited
SKU: AB263935
Price: 690 USD
Availability: InStock

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GPX4 KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9; X = 26 bp deletion, 2 bp insertion; Frameshift: 93.31%.

4 Images

Lab

Western blot - Human GPX4 knockout HeLa cell lysate (AB263935)

Lanes 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lanes 2 : GPX4 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lanes 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate, 20 ug

Lanes 1 - 3 : Merged signal (red and green). Green - ab125066 observed at 20 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab125066 was shown to react with Glutathione Peroxidase 4 in wild-type HeLa cells in Western blot with loss of signal observed in GPX4 knockout cell line ab262509 (knockout cell lysate ab263935). Wild-type HeLa and GPX4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab125066 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (<a href='/en-us/products/primary-antibodies/glutathione-peroxidase-4-antibody-epncir144-ab125066'>ab125066</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

GPX4 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human GPX4 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-gpx4-knockout-hela-cell-line-ab262509'>ab262509</a>)

Lane 3:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Predicted band size: 22 kDa

Observed band size: 20 kDa

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Lab

Western blot - Human GPX4 knockout HeLa cell lysate (AB263935)

ab206266 was shown to react with Glutathione Peroxidase 4 (HRP) in wild-type HeLa cells in western blot. Loss of signal was observed when GPX4 knockout cell line ab262509 (knockout cell lysate ab263935) was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab206266 overnight at 4 °C at a 1 in 5000 dilution Blots were developed with Optiblot ECL reagent (ab133456) and imaged.

All lanes:

Western blot - HRP Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (<a href='/en-us/products/primary-antibodies/hrp-glutathione-peroxidase-4-antibody-epncir144-ab206266'>ab206266</a>) at 1/5000 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

GPX4 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human GPX4 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-gpx4-knockout-hela-cell-line-ab262509'>ab262509</a>)

Lane 3:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 4:

Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 22 kDa

Observed band size: 20 kDa

false

Exposure time: 4min

Lab

Western blot - Human GPX4 knockout HeLa cell lysate (AB263935)

Lanes 1 - 3 : Merged signal (red and green). Green - ab41787 observed at 20 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab41787 was shown to react with Glutathione Peroxidase 4 in wild-type HeLa cells in Western blot with loss of signal observed in GPX4 knockout cell line ab262509 (knockout cell lysate ab263935). Wild-type HeLa and GPX4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab41787 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Glutathione Peroxidase 4 antibody (<a href='/en-us/products/primary-antibodies/glutathione-peroxidase-4-antibody-ab41787'>ab41787</a>) at 1 µg/mL

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

GPX4 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human GPX4 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-gpx4-knockout-hela-cell-line-ab262509'>ab262509</a>)

Lane 3:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Predicted band size: 22 kDa

Observed band size: 20 kDa

false

Supplier Data

Next Generation Sequencing - Human GPX4 knockout HeLa cell lysate (AB263935)

Knockout achieved by CRISPR/Cas9; X = 26 bp deletion, 2 bp insertion; Frameshift : 93.31%

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9; X = 26 bp deletion, 2 bp insertion; Frameshift: 93.31%

Disease

Adenocarcinoma

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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>" } } }

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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?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab263935-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human GPX4 knockout HeLa cell lysate", "number":"AB263935-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB263935-CMP01", "productcode":"" } ] } } }

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Properties and storage information

Gene name

GPX4

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Next Generation Sequencing, Western blot

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 Peroxidase 4 (GPX4) also known as PHGPx acts as an important enzyme in protecting cells from oxidative damage by reducing lipid hydroperoxides to their alcohols and free hydrogen peroxide to water. It has a mass of approximately 20 kDa. GPX4 is widely expressed in various tissues including the brain testis kidney and liver underlining its significance in different physiological processes. GPX4 is essential in cellular anti-oxidant defense making it a potential marker in studies using GPX4 western blot techniques.

Biological function summary

GPX4 plays an important role in protecting cellular membranes from lipid peroxidation by acting alone which differentiates it from other glutathione peroxidases that work in a complex. It is necessary for the production of glutathione products and helps maintain redox homeostasis by converting glutathione peroxides. This action prevents cell death pathways linked to oxidative stress and ensures cellular integrity particularly in sensitive tissues like the brain and reproductive organs.

Pathways

GPX4 integrates into important cellular antioxidant pathways. Specifically it functions within the glutathione metabolic pathway collaborating with related proteins like glutathione synthetase and glutathione reductase. GPX4 is also involved in ferroptosis regulation a cell death pathway critically linked to glutathione levels. It helps balance redox-sensitive signaling and metabolic activities demonstrating synergy with other antioxidant enzymes in the cellular response to oxidative stress.

Research connects GPX4 deficiency to neurological disorders and cancer. Its important antioxidative activity makes it significant in preventing neurodegenerative diseases such as Alzheimer's where it interacts with proteins like PINK1. GPX4 also shows relevance in cancer as its altered expression may increase vulnerability to cell death suggesting its potential role in chemotherapy resistance and therapy development.

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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

Visit the General protocols
Visit the Troubleshooting

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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'.

For licensing inquiries, please contact partnerships@abcam.com

Human GPX4 knockout HeLa cell lysate

Western blot - Human GPX4 knockout HeLa cell lysate (AB263935)

Western blot - Human GPX4 knockout HeLa cell lysate (AB263935)

Western blot - Human GPX4 knockout HeLa cell lysate (AB263935)

Next Generation Sequencing - Human GPX4 knockout HeLa cell lysate (AB263935)

Key facts

Cell type

Species or organism

Tissue

Knockout validation

Mutation description

Disease

Complementary Products

Cell Lines & Lysates

Primary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Product promise