Anti-Reduced Glutathione(GSH) antibody [EPR26828-37]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Reduced Glutathione(GSH) antibody [EPR26828-37] (AB326336)

Immunohistochemical analysis of paraffin-embedded Non-perfused fixed rat liver tissue labeling Glutathione with ab326336 at 1/200 (2.39 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on hepatocytes and plasma in rat liver.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Reduced Glutathione(GSH) antibody [EPR26828-37] (AB326336)

Immunohistochemical analysis of paraffin-embedded Non-perfused fixed mouse liver tissue labeling Glutathione with ab326336 at 1/200 (2.39 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on hepatocytes and plasma in mouse liver.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Reduced Glutathione(GSH) antibody [EPR26828-37] (AB326336)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A549 (human lung carcinoma epithelial cell) untreated A549 treated with 10µM BSO for 18 hours cells labelling Glutathione with ab326336 at 1/50 (9.56 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing decreased cytoplasmic staining in A549 cells (shown in green) treated with 10µM BSO for 18 hours. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Reduced Glutathione(GSH) antibody [EPR26828-37] (AB326336)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (fresh frozen) tissue labeling Glutathione with ab326336 at 1/50 (9.56 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 µg/mL dilution (Green).

Confocal image showing positive staining on mouse liver. The nuclear counterstain was DAPI (Blue). The section was incubated with the primary antibody for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 µg/mL dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Reduced Glutathione(GSH) antibody [EPR26828-37] (AB326336)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Glutathione with ab326336 at 1/100 (4.78 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on hepatocytes and plasma in human liver.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• I-ELISA

Lab

Indirect ELISA - Anti-Reduced Glutathione(GSH) antibody [EPR26828-37] (AB326336)

Indirect ELISA analysis of ab326336 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : Reduced Glutathione(GSH) cross-linked to BSA;Oxidized Glutathione(GSSG) cross-linked to BSA.

Antigen concentration : 1000 ng/ml

This antibody does not cross-react with Oxidized Glutathione(GSSG) by ELISA.

• IP

Lab

Immunoprecipitation - Anti-Reduced Glutathione(GSH) antibody [EPR26828-37] (AB326336)

Glutathione was immunoprecipitated from 0.35 mg Reduced Glutathione(GSH) cross-linked to BSA with ab326336 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab326336 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Reduced Glutathione(GSH) cross-linked to BSA
Lane 2 : ab326336 IP in Reduced Glutathione(GSH) cross-linked to BSA
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab326336 in reduced Glutathione(GSH) cross-linked to BSA whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 8 seconds.

All lanes:

Immunoprecipitation - Anti-Reduced Glutathione(GSH) antibody [EPR26828-37] (ab326336) at 1/1000 dilution

Lane 1:

Reduced Glutathione(GSH) cross-linked to BSA at 10 µg

Lane 2:

ab326336 at 1/30 IP in Reduced Glutathione(GSH) cross-linked to BSA

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab326336 in reduced Glutathione(GSH) cross-linked to BSA whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 75 kDa

false

Exposure time: 8s

• WB

Lab

Western blot - Anti-Reduced Glutathione(GSH) antibody [EPR26828-37] (AB326336)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with Oxidized Glutathione(GSSG) by western blot.

In Western blot, Anti-Albumin antibody [EPR12774] - Loading Control (ab192603) (1 : 1000) (67KDa).

All lanes:

Western blot - Anti-Reduced Glutathione(GSH) antibody [EPR26828-37] (ab326336) at 1/1000 dilution

Lane 1:

Reduced Glutathione(GSH) cross-linked to BSA at 10 ng

Lane 2:

Oxidized Glutathione(GSSG) cross-linked to BSA at 10 ng

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

Observed band size: 75 kDa,67 kDa

false

Exposure time: 59s