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    glutathione-peroxidase-1-antibody-epr3311-ab108429.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)

  • Datasheet
Reviews (1) Submit a question References (8)

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Western blot - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
  • Western blot - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
  • Flow Cytometry (Intracellular) - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
  • Western blot - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
  • Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3311] to Glutathione Peroxidase 1
  • Suitable for: Flow Cyt (Intra), WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-Glutathione Peroxidase 1 antibody [EPR3311]
    See all Glutathione Peroxidase 1 primary antibodies
  • Description

    Rabbit monoclonal [EPR3311] to Glutathione Peroxidase 1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • Human fetal liver, SH SY5Y, and THP1 cell lysates; Human breast carcinoma tissue. WB: HEK-293T cell lysate.
  • General notes

    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
    For more information see here.

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

    Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR3311
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Cancer
    • Cancer Metabolism
    • Cellular metabolic process
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Antioxidants
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • Anti-Glutathione Peroxidase 1 antibody [EPR3311] - BSA and Azide free (ab247669)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human Glutathione Peroxidase 1 protein (ab113603)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108429 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
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
1/1000 - 1/10000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.

Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
1/1000 - 1/10000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.

Target

  • Function

    Protects the hemoglobin in erythrocytes from oxidative breakdown.
  • Sequence similarities

    Belongs to the glutathione peroxidase family.
  • Cellular localization

    Cytoplasm.
  • Target information above from: UniProt accession P07203 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2876 Human
    • Entrez Gene: 24404 Rat
    • Omim: 138320 Human
    • SwissProt: P07203 Human
    • SwissProt: P04041 Rat
    • Unigene: 76686 Human
    • Unigene: 11323 Rat
    • Alternative names

      • AL033363 antibody
      • Cellular glutathione peroxidase antibody
      • Glutathione peroxidase 1 antibody
      • Glutathione peroxidase antibody
      • GPx 1 antibody
      • GPx-1 antibody
      • GPX1 antibody
      • GPX1_HUMAN antibody
      • GPXD antibody
      • GSHPx-1 antibody
      • GSHPX1 antibody
      • MGC14399 antibody
      • MGC88245 antibody
      see all

    Images

    • Western blot - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
      Western blot - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
      All lanes : Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : GPX1 knockout HEK-293T cell lysate

      Performed under reducing conditions.

      Predicted band size: 22 kDa
      Observed band size: 22 kDa



      False colour image of Western blot: Anti-Glutathione Peroxidase 1 antibody [EPR3311] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab108429 was shown to bind specifically to Glutathione Peroxidase 1. A band was observed at 22 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in GPX1 knockout cell line ab266650 (knockout cell lysate ab256932). To generate this image, wild-type and GPX1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    • Western blot - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
      Western blot - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: GPX1 knockout HAP1 cell lysate (20 µg)
      Lane 3: THP1 cell lysate (20 µg)
      Lane 4: HL60 cell lysate (20 µg)
      Lanes 1 and 2: Merged signal (red and green). Green - ab108429, observed at 22 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab108429 was shown to specifically react with Glutathione Peroxidase 1 in wild-type HAP1 cells. No band was observed when Glutathione Peroxidase 1 knockout samples were examined. Wild-type and Glutathione Peroxidase 1 knockout samples were subjected to SDS-PAGE. ab108429 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    • Flow Cytometry (Intracellular) - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
      Flow Cytometry (Intracellular) - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)

      Intracellular Flow Cytometry analysis of THP-1 (human acute monocytic leukemia) cells labeling Glutathione Peroxidase 1 with purified ab108429 at 1/250 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

       

       

    • Western blot - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
      Western blot - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
      All lanes : Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429) at 1/1000 dilution

      Lane 1 : Human fetal liver lysate
      Lane 2 : SH SY5Y cell lysate
      Lane 3 : THP1 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 22 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)

      ab108429, at 1/100 dilution, staining Glutathione Peroxidase 1 in paraffin-embedded Human breast carcinoma tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)
      Anti-Glutathione Peroxidase 1 antibody [EPR3311] (ab108429)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (8)

    Publishing research using ab108429? Please let us know so that we can cite the reference in this datasheet.

    ab108429 has been referenced in 8 publications.

    • Vindry C  et al. A Versatile Strategy to Reduce UGA-Selenocysteine Recoding Efficiency of the Ribosome Using CRISPR-Cas9-Viral-Like-Particles Targeting Selenocysteine-tRNA[Ser]Sec Gene. Cells 8:N/A (2019). PubMed: 31212706
    • Sonet J  et al. Selenized Plant Oil Is an Efficient Source of Selenium for Selenoprotein Biosynthesis in Human Cell Lines. Nutrients 11:N/A (2019). PubMed: 31277500
    • Jin L  et al. The PLAG1-GDH1 Axis Promotes Anoikis Resistance and Tumor Metastasis through CamKK2-AMPK Signaling in LKB1-Deficient Lung Cancer. Mol Cell 69:87-99.e7 (2018). PubMed: 29249655
    • Meimaridou E  et al. NNT is a key regulator of adrenal redox homeostasis and steroidogenesis in male mice. J Endocrinol 236:13-28 (2018). PubMed: 29046340
    • Sonet J  et al. Comparison of analytical methods using enzymatic activity, immunoaffinity and selenium-specific mass spectrometric detection for the quantitation of glutathione peroxidase 1. Anal Chim Acta 1011:11-19 (2018). PubMed: 29475480
    • Touat-Hamici Z  et al. Selenium-regulated hierarchy of human selenoproteome in cancerous and immortalized cells lines. Biochim Biophys Acta N/A:N/A (2018). Human . PubMed: 29660373
    • Chaisiriwong L  et al. A Case-Control Study of Involvement of Oxidative DNA Damage and Alteration of Antioxidant Defense System in Patients with Basal Cell Carcinoma: Modulation by Tumor Removal. Oxid Med Cell Longev 2016:5934024 (2016). IHC . PubMed: 27057281
    • Baqader NO  et al. Nuclear cytoplasmic trafficking of proteins is a major response of human fibroblasts to oxidative stress. J Proteome Res 13:4398-423 (2014). WB ; Human . PubMed: 25133973

    Customer reviews and Q&As

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    Flow Cytometry abreview for Anti-Glutathione Peroxidase 1 antibody [EPR3311]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Mouse Cell (Thymic Epithelial Cells)
    Permeabilization
    Yes - BD Permeabilization Buffer
    Gating Strategy
    EpCAM+ CD45- MHCII+ Zombie- cells
    Specification
    Thymic Epithelial Cells
    Preparation
    Cell harvesting/tissue preparation method: Digest of the thymus with Liberase and DNase in PBS
    Sample buffer: 1x PBS
    Fixation
    BD Fixation buffer containing Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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    Verified customer

    Submitted Jul 26 2022

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