Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
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
- Suitable for: Flow Cyt, WB, IHC-P, ICC
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Glutathione Peroxidase 4 antibody [EPNCIR144]
See all Glutathione Peroxidase 4 primary antibodies -
Description
Rabbit monoclonal [EPNCIR144] to Glutathione Peroxidase 4 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC HumanIHC-P HumanWB MouseHuman -
Immunogen
Synthetic peptide within Mouse Glutathione Peroxidase 4. The exact sequence is proprietary.
Database link: O70325 -
Positive control
- WB: Mouse testis, rat testis, human testis, human fetal liver, and human seminoma tissue lysates, LnCaP, Jurkat and HepG2 cell lysates. ICC/IF: HEK293 cells. IHC-P: Human kidney and stomach tissues. Flow Cyt: HeLa cells.
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General notes
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.
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).
See other anti-rabbit secondary antibodies that can be used with this 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 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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPNCIR144 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) at 1/20000 dilution (purified)
Lane 1 : Mouse testis lysate
Lane 2 : Rat testis lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 22 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST.
Dilution buffer: 5% NFDM/TBST. -
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.
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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.
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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.
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All lanes : Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) at 1/20000 dilution (purified)
Lane 1 : Human fetal liver tissue lysate
Lane 2 : Jurkat cell lysate
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 22 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST.
Dilution buffer: 5% NFDM/TBST. -
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.
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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.
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Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) at 1/5000 dilution (purified) + HepG2 cell lysate at 20 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 22 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST.
Dilution buffer: 5% NFDM/TBST. -
All lanes : Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) at 1/1000 dilution (unpurified)
Lane 1 : Human testis tissue lysate
Lane 2 : Human seminoma tissue lysate
Lane 3 : LnCaP cell lysate
Lane 4 : Human fetal liver tissue lysate
Lane 5 : Jurkat cell lysate
Lane 6 : HepG2 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-Rabbit HRP at 1/2000 dilution
Developed using the ECL technique.
Predicted band size: 22 kDa
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