Anti-GSH2 antibody [EPR29122-65] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSH2 antibody [EPR29122-65] - BSA and Azide free (AB321995)

This data was developed using ab321994, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse E12.5 brain tissue labeling GSH2 with ab321994 at 1/100 (5.19 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse E12.5 brain. The section was incubated with ab321994 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-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GSH2 antibody [EPR29122-65] - BSA and Azide free (AB321995)

This data was developed using ab321994, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse kidney (fresh frozen) tissue labeling GSH2 with ab321994 at 1/50 (10.38 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Negative control : confocal image showing no staining on mouse kidney. The nuclear counterstain was DAPI (Blue). The section was incubated with ab321994 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) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSH2 antibody [EPR29122-65] - BSA and Azide free (AB321995)

This data was developed using ab321994, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling GSH2 with ab321994 at 1/100 (5.19 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse cardiac muscle. The section was incubated with ab321994 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSH2 antibody [EPR29122-65] - BSA and Azide free (AB321995)

This data was developed using ab321994, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling GSH2 with ab321994 at 1/100 (5.19 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse kidney. The section was incubated with ab321994 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-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GSH2 antibody [EPR29122-65] - BSA and Azide free (AB321995)

This data was developed using ab321994, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse E14.5 cerebrum (fresh frozen) tissue labeling GSH2 with ab321994 at 1/50 (10.38 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Panel A : merged staining of anti-GSX2 (ab321994, green) and anti-Ki67 (ab279653, magenta) on mouse E14.5 cerebrum. GSH2 co-localize with Ki67 (PMID : 32234482).
Panel B : anti-GSX2 stained on mouse E14.5 cerebrum.
Panel C : anti-Ki67 stained in dividing cells on mouse E14.5 cerebrum.
The section was incubated with ab321994 and ab279653 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). 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) preadsorbedat 1/1000 2 ug/mL dilution.

• WB

Supplier Data

Western blot - Anti-GSH2 antibody [EPR29122-65] - BSA and Azide free (AB321995)

This data was developed using ab321994, the same antibody clone in a different buffer formulation.

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

Negative control : lung, heart, kidney.

GSX2 is specifically expressed in the LGE of the embryo brain( PMID : 7619729; PMID : 19709628; PMID : 32122989; PMID : 21205889)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-GSH2 antibody [EPR29122-65] (<a href='/en-us/products/primary-antibodies/gsh2-antibody-epr29122-65-ab321994'>ab321994</a>) at 1/1000 dilution

Lane 1:

Mouse E17 midbrain tissue lysate at 20 µg

Lane 2:

Mouse E17 kidney tissue lysate at 20 µg

Lane 3:

Mouse E17 brain tissue lysate at 20 µg

Lane 4:

Mouse E17 lung tissue lysate at 20 µg

Lane 5:

Mouse E14.5 heart tissue lysate at 20 µg

Lane 6:

Mouse E11.5 brain tissue lysate at 40 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 1/20000 dilution

Observed band size: 35 kDa,36 kDa

false

Exposure time: 180s

• Dot

Supplier Data

Dot Blot - Anti-GSH2 antibody [EPR29122-65] - BSA and Azide free (AB321995)

This data was developed using ab321994, the same antibody clone in a different buffer formulation.

Dot blot analysis of GSH2 using ab321994 at 1 : 1000 (0.519 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : 293T cells transfected with a mouse GSH2 expression vector containing a myc-His-tag® whole cell lysate
Lane2 : 293T cells transfected with a mouse GSH1 expression vector containing a myc-His-tag® whole cell lysate

Exposure time : 180 seconds.

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

This antibody does not cross-react with mouse GSH1.

#In Dot Blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Dot Blot - Anti-GSH2 antibody [EPR29122-65] (<a href='/en-us/products/primary-antibodies/gsh2-antibody-epr29122-65-ab321994'>ab321994</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with a mouse GSH2 expression vector containing a myc-His-tag&reg; whole cell lysate

Lane 2:

293T cells transfected with a mouse GSH1 expression vector containing a myc-His-tag&reg; whole cell lysate

Secondary

All lanes:

Dot 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

false

Exposure time: 180s