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AB262509

Human GPX4 knockout HeLa cell line

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(7 Publications)

GPX4 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 26 bp deletion, 2 bp insertion; Frameshift: 93.31%. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View Alternative Names

GPX4_HUMAN, GSHPx-4, Glutathione peroxidase 4, PHGPx, Phospholipid hydroperoxidase, Phospholipid hydroperoxide glutathione peroxidase, Phospholipid hydroperoxide glutathione peroxidase mitochondrial, Sperm nucleus glutathione peroxidase, mitochondrial, snGPx, snPHGPx

4 Images
Western blot - Human GPX4 knockout HeLa cell line (AB262509)
  • WB

Lab

Western blot - Human GPX4 knockout HeLa cell line (AB262509)

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 ° 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 (ab262509)

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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Western blot - Human GPX4 knockout HeLa cell line (AB262509)
  • WB

Lab

Western blot - Human GPX4 knockout HeLa cell line (AB262509)

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 sample was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab206266 overnight at 4 ° 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 (ab262509)

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

Western blot - Human GPX4 knockout HeLa cell line (AB262509)
  • WB

Lab

Western blot - Human GPX4 knockout HeLa cell line (AB262509)

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 ° 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 (ab262509)

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

Next Generation Sequencing - Human GPX4 knockout HeLa cell line (AB262509)
  • NGS

Supplier Data

Next Generation Sequencing - Human GPX4 knockout HeLa cell line (AB262509)

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

Form

Liquid

form

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

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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.

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What's included?

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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
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 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

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Target data

See full target information Phospholipid hydroperoxide glutathione peroxidase
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Publications (7)

Recent publications for all applications. Explore the full list and refine your search

Journal of nanobiotechnology 23:409 PubMed40457410

2025

Targeted tumor cell-intrinsic CTRP6 biomimetic codelivery synergistically amplifies ferroptosis and immune activation to boost anti-PD-L1 immunotherapy efficacy in lung cancer.

Applications

Unspecified application

Species

Unspecified reactive species

Songhua Cai,Jing Huang,Hongjie Fan,Zhilin Sui,Chujian Huang,Youjun Deng,Ran Jia,Lixu Wang,Kai Ma,Xiaotong Guo,Jie He,Baihua Zhang,Zhentao Yu

Translational cancer research 14:1311-1322 PubMed40104701

2025

Total Astragalus saponins promote ferroptosis in gastric cancer cells by upregulating SIRT3.

Applications

Unspecified application

Species

Unspecified reactive species

Yue Zou,Jingling Zhao,Chengyin Li,Rui Wang,Xiaocui Jiang,Zhongyi Zhu,Qiyuan Wang,Min Xiao

Journal of translational medicine 23:250 PubMed40022084

2025

Hsa_circ_0002301 inhibits ferroptosis in gastric cancer by encoding the de novo protein HECTD1-463aa.

Applications

Unspecified application

Species

Unspecified reactive species

Song Wang,Chengwei Wu,Jiawei Wang,Feng Yuan,Yinfen Hou,Tingting Cao,Lishuai Xu,Long Qian,Yabin Xia,Li Xu,Ailiang Zeng,Xiaoming Wang,Luman Wang,Xiaoxu Huang

Cell death & disease 15:79 PubMed38246916

2024

CYLD regulates cell ferroptosis through Hippo/YAP signaling in prostate cancer progression.

Applications

Unspecified application

Species

Unspecified reactive species

Yanan Gu,Shiqi Wu,Junjie Fan,Zeji Meng,Guoqiang Gao,Tianjie Liu,Qi Wang,Huayu Xia,Xinyang Wang,Kaijie Wu

Heliyon 9:e17849 PubMed37501954

2023

Effect and mechanism of circHMGA2 on ferroptosis and pyroptosis in myocardial ischemia-reperfusion model CircHMGA2 exacerbates MI/R injury.

Applications

Unspecified application

Species

Unspecified reactive species

Pin Feng,Yi Chu,Jun Li,Jingyi Dang,Jianghong Chen,Wei Zhang

Journal of orthopaedic surgery and research 18:518 PubMed37480032

2023

Circ-STC2 promotes the ferroptosis of nucleus pulposus cells via targeting miR-486-3p/TFR2 axis.

Applications

Unspecified application

Species

Unspecified reactive species

Liangping Xiong,Xiaoyan Li,Xi Hua,Zhonglai Qian

Frontiers in pharmacology 13:869300 PubMed35517804

2022

Activation of the PPARγ Prevents Ferroptosis-Induced Neuronal Loss in Response to Intracerebral Hemorrhage Through Synergistic Actions With the Nrf2.

Applications

Unspecified application

Species

Unspecified reactive species

Chenyang Duan,Dian Jiao,Hanbin Wang,Qiaoli Wu,Weidong Men,Hua Yan,Chunhui Li
View all publications
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