Anti-GPT2 antibody [EPR29176-18] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPT2 antibody [EPR29176-18] - BSA and Azide free (AB324833)

This data was developed using ab322261, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling GPT2 with ab322261 at 1/2000 (0.261 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human kidney.
The section was incubated with ab322261 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-GPT2 antibody [EPR29176-18] - BSA and Azide free (AB324833)

This data was developed using ab322261, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling GPT2 with ab322261 at 1/2000 (0.261 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : No staining on human cerebrum (PMID : 17826732).
The section was incubated with ab322261 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-GPT2 antibody [EPR29176-18] - BSA and Azide free (AB324833)

This data was developed using ab322261, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human cardiac muscle staining of GPT2 with ab322261 at a 1/2000 (0.261 µg/ml) dilution and COX IV with ab202554 at 1/500 (0.228 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-GPT2 (green; Opal™570) and anti-COX IV (magenta; Opal™520) on human cardiac muscle.
Panel B : anti-GPT2 showed granular staining in human cardiac muscle.
Panel C : ant-COX IV showed granular staining in human cardiac muscle.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322261 and ab202554 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-GPT2 antibody [EPR29176-18] - BSA and Azide free (AB324833)

This data was developed using ab322261, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human kidney staining of GPT2 with ab322261 at a 1/2000 (0.261 µg/ml) dilution and COX IV with ab202554 at 1/500 (0.228 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-GPT2 (green; Opal™570) and anti-COX IV (magenta; Opal™520) on human kidney.
Panel B : anti-GPT2 showed granular staining in human kidney.
Panel C : ant-COX IV showed granular staining in human kidney.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322261 and ab202554 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GPT2 antibody [EPR29176-18] - BSA and Azide free (AB324833)

This data was developed using ab322261, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling GPT2 with ab322261 at 1/500 (1.044 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing mitochondrial staining in HepG2 cells (shown in green). 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).

anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/500 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab322261 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody preadsorbed at 1/1000 2 ug/ml dilution -ve control 2 : anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/500 5 ug/ml dilution , followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPT2 antibody [EPR29176-18] - BSA and Azide free (AB324833)

This data was developed using ab322261, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling GPT2 with ab322261 at 1/2000 (0.261 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human cardiac muscle (PMID : 17826732, PMID : 19360321).
The section was incubated with ab322261 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-GPT2 antibody [EPR29176-18] - BSA and Azide free (AB324833)

This data was developed using ab322261, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling GPT2 with ab322261 at 1/2000 (0.261 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human liver.
The section was incubated with ab322261 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

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GPT2 antibody [EPR29176-18] - BSA and Azide free (AB324833)

This data was developed using ab322261, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized C2C12 (mouse myoblast) cells labelling GPT2 with ab322261 at 1/500 (1.044 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing mitochondrial staining in C2C12 cells (shown in green). 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).

anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/500 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab322261 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody preadsorbed at 1/1000 2 ug/ml dilution -ve control 2 : anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/500 5 ug/ml dilution , followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-GPT2 antibody [EPR29176-18] - BSA and Azide free (AB324833)

This data was developed using ab322261, the same antibody clone in a different buffer formulation.

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

In lane 3, this blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

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

Exposure time : Lanes 1-2 : 37 seconds, Lane 3 : 114 seconds

All lanes:

Western blot - Anti-GPT2 antibody [EPR29176-18] (<a href='/en-us/products/primary-antibodies/gpt2-antibody-epr29176-18-ab322261'>ab322261</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocelullar carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HepG2 transfected with siRNA specifically targeting GPT2 whole cell lysate at 20 µg

Lane 3:

HT-29 (human colorectal adenocarcinoma 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

Observed band size: 58 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-GPT2 antibody [EPR29176-18] - BSA and Azide free (AB324833)

This data was developed using ab322261, the same antibody clone in a different buffer formulation.

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

The identity of the higher MW bands in lanes 2-4 are unknown.

All lanes:

Western blot - Anti-GPT2 antibody [EPR29176-18] (<a href='/en-us/products/primary-antibodies/gpt2-antibody-epr29176-18-ab322261'>ab322261</a>) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate at 20 µg

Lane 2:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 3:

Mouse heart tissue lysate at 20 µg

Lane 4:

Human heart tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 46 kDa

false

Exposure time: 136s

• WB

Supplier Data

Western blot - Anti-GPT2 antibody [EPR29176-18] - BSA and Azide free (AB324833)

This data was developed using ab322261, the same antibody clone in a different buffer formulation.

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

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

Exposure time : Lanes 1-2 : 8 seconds, Lanes 3-4 : 10 seconds

All lanes:

Western blot - Anti-GPT2 antibody [EPR29176-18] (<a href='/en-us/products/primary-antibodies/gpt2-antibody-epr29176-18-ab322261'>ab322261</a>) at 1/1000 dilution

Lane 1:

HEK-293 (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HEK-293 transfected with siRNA specifically targeting GPT2 whole cell lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

C2C12 (mouse myoblast) 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 1/20000 dilution

Observed band size: 58 kDa,36 kDa

false

• Dot

Supplier Data

Dot Blot - Anti-GPT2 antibody [EPR29176-18] - BSA and Azide free (AB324833)

This data was developed using ab322261, the same antibody clone in a different buffer formulation.

Dot blot analysis of GPT2 using ab322261 at 1 : 1000 (0.522 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : His-tagged human GPT1 fragment
Lane2 : His-tagged human GPT2 fragment

Exposure time : 180 seconds.

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

This antibody does not cross-react with human GPT1.

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

All lanes:

Dot Blot - Anti-GPT2 antibody [EPR29176-18] (<a href='/en-us/products/primary-antibodies/gpt2-antibody-epr29176-18-ab322261'>ab322261</a>) at 1/1000 dilution

Lane 1:

His-tagged human GPT1 fragment

Lane 2:

His-tagged human GPT2 fragment

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