Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-GST3 / GST pi antibody [EPR20554] (ab241331)

Overview

  • Product name

    Anti-GST3 / GST pi antibody [EPR20554]
    See all GST3 / GST pi primary antibodies
  • Description

    Rabbit monoclonal [EPR20554] to GST3 / GST pi
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant full length protein within Human GST3/ GST pi aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P09211

  • Positive control

    • WB: Wild-type HAP1, PC-3, Jurkat and HEK-293 whole cell lysates; Human brain and heart lysates. IHC-P: Human pancreas, tonsil and kidney tissues. ICC/IF: Wild-type HAP1 and Jurkat cells. Flow Cyt: Wild-type HAP1 cells.
  • General notes

    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 ab241331 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
WB 1/1000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/250.
Flow Cyt 1/300.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Conjugation of reduced glutathione to a wide number of exogenous and endogenous hydrophobic electrophiles.
    • Sequence similarities

      Belongs to the GST superfamily. Pi family.
      Contains 1 GST C-terminal domain.
      Contains 1 GST N-terminal domain.
    • Information by UniProt
    • Database links

    • Alternative names

      • Deafness antibody
      • Deafness X-linked 7 antibody
      • DFN7 antibody
      • FAEES3 antibody
      • Fatty Acid Ethyl Ester Synthase III antibody
      • Glutathione S Transferase 3 antibody
      • Glutathione S Transferase Pi antibody
      • Glutathione S-transferase P antibody
      • Glutathione S-transferase pi 1 antibody
      • GST class-pi antibody
      • GST3 antibody
      • GSTP antibody
      • Gstp1 antibody
      • GSTP1-1 antibody
      • GSTP1_HUMAN antibody
      • PI antibody
      • X linked 7 antibody
      see all

    Images

    • All lanes : Anti-GST3 / GST pi antibody [EPR20554] (ab241331) at 1/1000 dilution

      Lane 1 : Human brain tissue lysate
      Lane 2 : Human heart tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/100000 dilution

      Predicted band size: 23 kDa
      Observed band size: 23 kDa


      Exposure time: 52 seconds


      Blocking/Diluting buffer and concentration: 5% NFDM/TBST

    • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling GST3 / GST pi with ab241331 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining on tubules of human kidney (PMID: 1977319). Counterstained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

       

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling GST3 / GST pi with ab241331 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining on subset immune cells of human tonsil. (PMID: 18604726). Counterstained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

       

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling GST3 / GST pi with ab241331 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining on duct epithelium of human pancreas. (PMID: 1977319). Counterstained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

       

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (human T cell leukemia T lymphocyte) cells labeling GST3 GST pi (Green) with ab241331 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. Confocal image showing cytoplasmic staining in Jurkat cell line.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized wild type and GST pi KO HAP1 (GST pi knockout human chronic myelogenous leukemia near-haploid cell line) cells labeling GST3 / GST pi (Green) with ab241331 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. Confocal image showing cytoplasmic staining in parental HAP1 cell line and no staining in GST pi KO HAP1 cell line.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized GST pi KO HAP1 (human chronic myelogenous leukemia near-haploid cell line, Left) / Wild-type HAP1 (Right) cell line labeling GST3 / GST pi with ab241331 at 1/300 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    • All lanes : Anti-GST3 / GST pi antibody [EPR20554] (ab241331) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 cell lysate
      Lane 2 : GST3 / GST pi knockout HAP1 cell lysate
      Lane 3 : PC-3 (human prostate adenocarcinoma epithelial cell), whole cell lysate
      Lane 4 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
      Lane 5 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Developed using the ECL technique.

      Predicted band size: 23 kDa
      Observed band size: 23 kDa


      Exposure time: 15 seconds


      Blocking/Diluting buffer and concentration: 5% NFDM/TBST 

      ab241331 was shown to specifically react with GST3 / GST pi in wild-type HAP1 cells as signal was lost in GST3 / GST pi knockout cells. Wild-type and GST3 / GST pi knockout samples were subjected to SDS-PAGE. ab241331 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

    References

    ab241331 has not yet been referenced specifically in any publications.

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