Recombinant Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] - BSA and Azide free (ab219592)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPNCIR144] to Glutathione Peroxidase 4 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] - BSA and Azide free
See all Glutathione Peroxidase 4 primary antibodies -
Description
Rabbit monoclonal [EPNCIR144] to Glutathione Peroxidase 4 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, Jurkat cell lysate; HepG2, A549, 293T whole cell lysate; rat heart lysate, mouse ovary, heart and lung lysate; rat, human and mouse testis tissue lysates. ICC/IF: HEK293 and HeLa cells. IHC-P: Human kidney and stomach tissues. Flow Cyt (intra): HeLa cells.
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General notes
ab219592 is the carrier-free version of ab125066.
This antibody was developed as part of a collaboration between Epitomics, the National Cancer Institute's Center for Cancer Research and the lab of Dolph Hatfield. View antibodies from NCI Center for Cancer Research Collaboration.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPNCIR144 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab219592 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 17 kDa (predicted molecular weight: 22 kDa).
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Detects a band of approximately 17 kDa (predicted molecular weight: 22 kDa). |
Target
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Function
Protects cells against membrane lipid peroxidation and cell death. Required for normal sperm development and male fertility. Could play a major role in protecting mammals from the toxicity of ingested lipid hydroperoxides. Essential for embryonic development. Protects from radiation and oxidative damage. -
Tissue specificity
Present primarily in testis. -
Sequence similarities
Belongs to the glutathione peroxidase family. -
Cellular localization
Mitochondrion. Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 2879 Human
- Entrez Gene: 625249 Mouse
- Entrez Gene: 29328 Rat
- Omim: 138322 Human
- SwissProt: P36969 Human
- SwissProt: O70325 Mouse
- SwissProt: P36970 Rat
- Unigene: 433951 Human
see all -
Alternative names
- Glutathione peroxidase 4 antibody
- GPX 4 antibody
- GPX-4 antibody
see all
Images
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All lanes : 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
Lane 2 : GPX4 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 22 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab125066).
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.
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All lanes : Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) at 1/1000 dilution
Lane 1 : 293T (Human embryonic kidney epithelial cell) whole cell lysate
Lane 2 : A549 (Human lung carcinoma epithelial cell) whole cell lysate
Lane 3 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 4 : Rat heart lysate
Lane 5 : Mouse ovary lysate
Lane 6 : Mouse heart lysate
Lane 7 : Mouse lung lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 22 kDa
Observed band size: 22 kDa
Exposure time: 20 secondsThis data was developed using the same antibody clone in a different buffer formulation (ab125066).
Blocking/Diluting buffer and concentration 5% NFDM/TBST.
ab181602 was used as GAPDH loading control.
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Unpurified ab125066, at a 1/100 dilution, staining Glutathione Peroxidase 4 in paraffin-embedded Human kidney tissue by immunohistochemistry.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125066).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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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 Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125066).
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All lanes : Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) at 1/1000 dilution
Lane 1 : Rat testis tissue lysate
Lane 2 : Mouse testis tissue lysate
Lane 3 : Human testis tissue lysate
Lysates/proteins at 20 µg per lane.
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: 19,22 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Diluting buffer and concentration: 5% NFDM/TBST
This antibody can detect 19kda cytoplasmic form and 22kda mitochondrial isoform.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125066).
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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 (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with 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 (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125066).
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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 in PBS) for 1 hour at 22°C. ab150081 an Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Nuclear staining was carried out with DAPI.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125066).
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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 (ab97051) at 1/500. 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125066).
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (4)
ab219592 has been referenced in 4 publications.
- Dang G et al. T lymphocyte-derived extracellular vesicles aggravate abdominal aortic aneurysm by promoting macrophage lipid peroxidation and migration via pyruvate kinase muscle isozyme 2. Redox Biol 50:102257 (2022). PubMed: 35149342
- Zhang S et al. Targeting ferroptosis with miR-144-3p to attenuate pancreatic β cells dysfunction via regulating USP22/SIRT1 in type 2 diabetes. Diabetol Metab Syndr 14:89 (2022). PubMed: 35761309
- Zhou Y The Protective Effects of Cryptochlorogenic Acid on ß-Cells Function in Diabetes in vivo and vitro via Inhibition of Ferroptosis. Diabetes Metab Syndr Obes 13:1921-1931 (2020). PubMed: 32606852
- Wu C et al. Induction of ferroptosis and mitochondrial dysfunction by oxidative stress in PC12 cells. Sci Rep 8:574 (2018). PubMed: 29330409