Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)

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Overview

  • Product name

    Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free
    See all GST3 / GST pi primary antibodies

  • Description

    Rabbit monoclonal [EPR20554] to GST3 / GST pi - BSA and Azide free

  • 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. IHC-P: Human pancreas, tonsil and kidney tissues. ICC/IF: Wild-type HAP1 and Jurkat cells. Flow Cyt: Wild-type HAP1 cells.

  • General notes

    Ab242014 is the carrier-free version of ab241331. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    ab242014 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 ab242014 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 Use at an assay dependent concentration. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
  • 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

      • Western blot - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)
        Western blot - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)
        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

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



        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.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241331).

         

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)

        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.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241331).

         

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

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)

        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.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241331).

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

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)

        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.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241331).

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

      • Immunocytochemistry/ Immunofluorescence - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)
        Immunocytochemistry/ Immunofluorescence - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)

        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.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241331).

      • Immunocytochemistry/ Immunofluorescence - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)
        Immunocytochemistry/ Immunofluorescence - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)

        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.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241331).

      • Flow Cytometry - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)
        Flow Cytometry - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (ab242014)

        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.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241331).

      References

      ab242014 has not yet been referenced specifically in any publications.

      Publishing research using ab242014? Please let us know so that we can cite the reference in this datasheet.

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