Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free

• IHC-P

Supplier Data

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.

• ICC/IF

Supplier Data

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 Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (AB242014)

Intracellular 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).

• IHC-P

Supplier Data

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.

• ICC/IF

Supplier Data

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).

• IHC-P

Supplier Data

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.

• WB

Supplier Data

Western blot - Anti-GST3 / GST pi antibody [EPR20554] - BSA and Azide free (AB242014)

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).

All lanes:

Western blot - Anti-GST3 / GST pi antibody [EPR20554] (<a href='/en-us/products/primary-antibodies/gst3-gst-pi-antibody-epr20554-ab241331'>ab241331</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

GST3 / GST pi knockout HAP1 cell lysate at 20 µg

Lane 3:

PC-3 (human prostate adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 5:

HEK-293 (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 23 kDa

Observed band size: 23 kDa

true

Exposure time: 15s