Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Glutathione Peroxidase 1 antibody [EPR3312] (HRP) (ab197034)

Overview

  • Product name

    Anti-Glutathione Peroxidase 1 antibody [EPR3312] (HRP)
    See all Glutathione Peroxidase 1 primary antibodies
  • Description

    Rabbit monoclonal [EPR3312] to Glutathione Peroxidase 1 (HRP)
  • Host species

    Rabbit
  • Conjugation

    HRP
  • Tested applications

    Suitable for: IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Common marmoset
  • Immunogen

    Synthetic peptide. within Human Glutathione Peroxidase 1 aa 150 to the C-terminus (internal sequence). The exact sequence is proprietary.
    Database link: P07203
    (Peptide available as ab156307)

  • Positive control

    • WB: THP1 whole cell lysate.. IHC-P: FFPE human normal colon tissue sections.
  • General notes

    Alternative versions available:
    Anti-Glutathione Peroxidase 1 antibody [EPR3312] (ab108427) - Knockout validated

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab197034 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
IHC-P 1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/5000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).Can be blocked with Glutathione Peroxidase 1 peptide (ab156307).

Target

Images

  • All lanes : Anti-Glutathione Peroxidase 1 antibody [EPR3312] (HRP) (ab197034) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : GPX1 (Glutathione Peroxidase 1) knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 22 kDa


    Exposure time: 3 minutes


    ab197034 was shown to recognize Glutathione Peroxidase 1 in wild-type HAP1 cells as signal was lost at the expected MW in GPX1 (Glutathione Peroxidase 1) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GPX1 (Glutathione Peroxidase 1) knockout samples were subjected to SDS-PAGE. Ab197034 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

  • Anti-Glutathione Peroxidase 1 antibody [EPR3312] (HRP) (ab197034) at 1/5000 dilution + THP1 (Human acute monocytic leukemia cell line) Whole Cell Lysate at 10 µg

    Developed using the ECL technique.

    Performed under reducing conditions.

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


    Exposure time: 20 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab197034 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

  • IHC image of Glutathione Peroxidase 1 staining in a section of formalin-fixed paraffin-embedded human normal colon*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab197034, at a dilution of 1/50, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

     

References

ab197034 has not yet been referenced specifically in any publications.

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