Name: Anti-Glutathione Peroxidase 4 antibody [EPNCIR144]
SKU: ab125066
Rating: 5 (13 reviews)

Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] is a rabbit recombinant monoclonal antibody that is used to detect Glutathione Peroxidase 4 in Flow cytometry (Intra), ICC/IF, IHC-P, Western blot. Suitable for Human, Mouse, Rat samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Antibody clone EPNCIR144 is the most widely used clone for Glutathione Peroxidase 4 on the market
- Specificity confirmed with GPX4 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (AB125066), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	ICC/IF	WB	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested
Mouse	Expected	Expected	Tested	Expected
Rat	Expected	Expected	Tested	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50.00000 - 1/250.00000	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000.00000 - 1/10000.00000	Notes -
Species Rat	Dilution info 1/1000.00000 - 1/10000.00000	Notes -
Species Human	Dilution info 1/1000.00000 - 1/10000.00000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/400	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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Target data

See full target information Phospholipid hydroperoxide glutathione peroxidase

Function

Essential antioxidant peroxidase that directly reduces phospholipid hydroperoxide even if they are incorporated in membranes and lipoproteins (By similarity). Can also reduce cholesterol hydroperoxide and thymine hydroperoxide (By similarity). Plays a key role in protecting cells from oxidative damage by preventing membrane lipid peroxidation (By similarity). Required to prevent cells from ferroptosis, a non-apoptotic cell death resulting from an iron-dependent accumulation of lipid reactive oxygen species (PubMed:24439385). The presence of selenocysteine (Sec) versus Cys at the active site is essential for life: it provides resistance to overoxidation and prevents cells against ferroptosis (By similarity). The presence of Sec at the active site is also essential for the survival of a specific type of parvalbumin-positive interneurons, thereby preventing against fatal epileptic seizures (By similarity). May be required to protect cells from the toxicity of ingested lipid hydroperoxides (By similarity). Required for normal sperm development and male fertility (By similarity). Essential for maturation and survival of photoreceptor cells (By similarity). Plays a role in a primary T-cell response to viral and parasitic infection by protecting T-cells from ferroptosis and by supporting T-cell expansion (By similarity). Plays a role of glutathione peroxidase in platelets in the arachidonic acid metabolism (PubMed:11115402). Reduces hydroperoxy ester lipids formed by a 15-lipoxygenase that may play a role as down-regulator of the cellular 15-lipoxygenase pathway (By similarity). Can reduce fatty acid-derived hydroperoxides (PubMed:11115402, PubMed:36608588). Can also reduce small soluble hydroperoxides such as H2O2, cumene hydroperoxide and tert-butyl hydroperoxide (PubMed:17630701, PubMed:36608588).

Alternative names

Phospholipid hydroperoxide glutathione peroxidase GPX4, PHGPx, Glutathione peroxidase 4, GPx-4, GSHPx-4, GPX4

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPNCIR144
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Stable for 12 months at -20°C

Notes

Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-P and WB.

Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) was first used in a scientific publication in 2012 and has been cited over 567 times in peer reviewed journals. It's performance in Western Blot and IHC in mouse and human samples is trusted by the scientific community.

Abcam's high quality manufacturing and validation processes ensure Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

The specificity of Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) has been confirmed by Western Blot testing in Glutathione Peroxidase 4 knockout HeLa cells (Human GPX4 knockout HeLa cell line ab262509).

Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) has 12 independent reviews from customers.

Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) specifically detects Glutathione Peroxidase 4 (UniProt ID: O70325; Molecular weight: 22kDa) and is sold in 100 µL and 1 mL selling sizes.

Conjugation-ready, carrier free format available for antibody clone EPNCIR144 - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] - BSA and Azide free ab219592.

Antibody clone EPNCIR144 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor^® 488, Alexa Fluor^® 647 (Alexa Fluor® 488 Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] ab314333, Alexa Fluor® 647 Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] ab314334).

Top cited antibody for this target with >1000 citations. Glutathione peroxidase 4 is a master regulator of ferroptosis and lipid peroxidation. This antibody is crucial in cancer research, particularly in understanding GPX4's role in cell death and antioxidant defence and is widely used in studies of reactive oxygen species (ROS) and GPX4 inhibitors for therapeutic applications.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

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

Activity summary

Glutathione Peroxidase 4 (GPX4) also known as GXP4 or 1B4 is an essential selenoprotein with a molecular weight of approximately 22 kDa. It acts mechanically by reducing lipid hydroperoxides to their corresponding alcohols and free hydrogen peroxide to water utilizing glutathione as a cofactor. GPX4 is ubiquitously expressed with high expression levels found in the testis indicating its importance in spermatogenesis.

Biological function summary

This enzyme protects cells from oxidative damage by maintaining cellular redox balance. GPX4 exists as a monomer and is not part of larger complexes. It enables cells to counteract the harmful effects of reactive oxygen species (ROS) safeguarding lipid membranes from peroxidation. The protective function extends to the nervous system and other tissues where oxidative stress might lead to cellular damage.

Pathways

This enzyme plays a critical role in the antioxidant defense system. It integrates with the glutathione pathway where it operates alongside glutathione reductase and glutathione synthetase to detoxify cells. The protein also interacts with lipid metabolism pathways maintaining membrane integrity and function. These pathways further link GPX4 with other oxidative stress-related proteins and cellular survival mechanisms.

Associated diseases and disorders

GPX4 has connections to neurodegenerative diseases like Parkinson's disease and cancer. In neurodegeneration low GPX4 levels can lead to increased lipid peroxidation highlighting its protective function against neuronal damage. In cancer altered GPX4 expression associates with the oxidative stress response affecting tumor progression. The protein's interactions with key oxidative stress regulators suggest potential targets for therapeutic intervention.

Product promise

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12 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Glutathione Peroxidase 4 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human testis using rabbit Anti-Glutathione Peroxidase 4 antibody
Immunohistochemical analysis of formalin fixed paraffin embedded human testis labelling Glutathione Peroxidase 4 with ab125066 at a concentration of 0.1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). ab125066 anti-Glutathione Peroxidase 4 antibody [EPNCIR144] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
This image is courtesy of a customer review submitted by Kirk McManus
Immunocytochemistry/ Immunofluorescence - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Glutathione Peroxidase 4 Immunocytochemistry/ Immunofluorescence staining of HeLa cells using rabbit Anti-Glutathione Peroxidase 4 antibody
Unpurified ab125066 staining Glutathione Peroxidase 4 in the HeLa cell line from Human cervical cancer by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methanol. Samples were incubated with primary antibody (1/500 dilution in PBS) for 1 hour at 22°C. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 an Alexa Fluor^® 488-conjugated Goat anti-rabbit IgG polyclonal (1/200 dilution) was used as the secondary antibody. Nuclear staining was carried out with DAPI.
Western blot - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Lanes 1 - 3: Merged signal (red and green). Green - ab125066 observed at 20 kDa. Red - loading control Anti-GAPDH antibody [6C5] - 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 Human GPX4 knockout HeLa cell line ab262509 (knockout cell lysate Human GPX4 knockout HeLa 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 Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed 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] (ab125066) 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 (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
Performed under reducing conditions.
Predicted band size: 22 kDa
Observed band size: 20 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Glutathione Peroxidase 4 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human kidney tissue using rabbit Anti-Glutathione Peroxidase 4 antibody
Unpurified ab125066 at a 1/100 dilution staining Glutathione Peroxidase 4 in paraffin-embedded human kidney tissue by immunohistochemistry.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Glutathione Peroxidase 4 Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-Glutathione Peroxidase 4 antibody
Immunofluorescence staining of HEK293 cells with purified ab125066 at a working dilution of 1/200 counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) used at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 a mouse anti-tubulin antibody (1/1000) was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (Alexa Fluor^® 594 goat anti-mouse 1/1000) shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1 purified ab125066 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at a dilution of 1/500. For negative control 2 Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1/400.
Flow Cytometry (Intracellular) - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Glutathione Peroxidase 4 Flow Cytometry (Intracellular) staining of HeLa (human cervix adenocarcinoma) cells using rabbit Anti-Glutathione Peroxidase 4 antibody
Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Glutathione Peroxidase 4 (red) with ab125066 at a 1/400 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
Western blot - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Blocking buffer: 5% NFDM/TBST.
Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) at 1/20000 dilution
Lane 1: Mouse testis lysate at 20 µg
Lane 2: Rat testis lysate at 20 µg
Secondary
All lanes: HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 22 kDa
Observed band size: 17 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Glutathione Peroxidase 4 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human stomach tissue using rabbit Anti-Glutathione Peroxidase 4 antibody
Immunohistochemical staining of paraffin embedded human stomach with purified ab125066 at a working dilution of 1/50. The secondary antibody used is HRP goat anti-rabbit IgG H&L (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
Western blot - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Blocking buffer and concentration: 5% NFDM/TBST

Diluting buffer and concentration: 5% NFDM/TBST

This antibody can detect 19kda cytoplasmic form and 22kda mitochondrial isoform.
All lanes: Western blot - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) at 1/1000 dilution
Lane 1: Rat testis tissue lysate at 20 µg
Lane 2: Mouse testis tissue lysate at 20 µg
Lane 3: Human testis tissue lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 22 kDa
Observed band size: 19 kDa, 22 kDa
Exposure time: 1s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Glutathione Peroxidase 4 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human testis using rabbit Anti-Glutathione Peroxidase 4 antibody
Immunohistochemical analysis of formalin fixed paraffin embedded human testis labelling Glutathione Peroxidase 4with ab125066 at a concentration of 1 µg/ml. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins. ab125066 anti-Glutathione Peroxidase 4 antibody [EPNCIR144] was incubated for 30 mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.
Biochemical assay - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Niu et al used Lipid Peroxidation (MDA) Assay Kit Lipid Peroxidation (MDA) Assay Kit (Colorimetric/Fluorometric) ab118970, GSH/ GSSG Assay Kit GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) ab138881, and Glutathione Peroxidase Assay Kit ab125066 to investigate the use of porous Se@SiO2 nanospheres to achieve controlled release of selenium, a scavenger of intracellular free radicals, in mouse eyes.
Male diabetic db/db and control db/m mice were injected in the eyes with porous Se@SiO2 nanospheres, and porous SiO2 nanospheres (NPs) without Se.
Porous Se@SiO2 nanospheres inhibit diabetes-induced retinal lipid peroxidation and inflammation. Levels of MDA in retinal homogenates (n = 6). Expression levels of GPX4 in retinas were measured using western blotting; β‐actin was used as a loading control (left panel). Band densities were assessed using the ImageJ software, and GPX4 expression levels are represented as their ratios to β‐actin (right panel) (n = 3). Levels of GSH, GSSG, and the ratio of GSH to GSSG in retinal homogenates (n = 6). Data are represented as the mean ± SD. **p < 0.01, ***p < 0.001; ns, nonsignificant.
Retinal samples were harvested, washed, and lysed according to the manufacturer’s instructions. Protein concentrations were determined using a bicinchoninic acid kit. MDA and glutathione concentrations were determined using a lipid peroxidation MDA assay kit, reduced glutathione (GSH) / oxidized glutathione (GSSG) Ratio Detection Assay kit (Lipid Peroxidation (MDA) Assay Kit (Colorimetric/Fluorometric) ab118970, GSH/GSSG Ratio Detection Assay Kit (Fluorometric - Green) ab138881; Abcam, MA, USA), and a microplate reader. The relative concentrations of MDA and glutathione were calculated by normalizing the measured concentrations to that of the total protein.
Western blot - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Blocking/Diluting buffer and concentration 5% NFDM/TBST.

Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as GAPDH loading control.
All lanes: Western blot - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) at 1/1000 dilution
Lane 1: 293T (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 2: A549 (Human lung carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 3: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4: Rat heart lysate at 20 µg
Lane 5: Mouse ovary lysate at 20 µg
Lane 6: Mouse heart lysate at 20 µg
Lane 7: Mouse lung lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 22 kDa
Observed band size: 22 kDa
Exposure time: 20s