Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Adenine Nucleotide Translocator 2/ANT 2 with ab325120 at 1/50 (10.0 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in HeLa cell line, which is co-staining with mitochondrial marker (shown in magenta). The counterstain was observed in green. 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 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab325120 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution.
-ve control 2 : anti-COX IV mouse monoclonal antibody - Mitochondrial Marker at 1/200 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining Adenine Nucleotide Translocator 2/ANT 2 with ab325120 at a 1 : 100 (5.04 ug/ml) dilution, ab202554 anti-COX IV used at 1 : 500 (0.23 ug/ml) dilution.

Panel A : merged staining of anti-ANT 2 (green; Opal™520) and anti-COX IV (magenta; Opal™570) on human liver.
Panel B : anti-ANT 2 showed mitochondrial staining in human liver.
Panel C : anti-COX IV showed mitochondrial staining in human liver.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325120 and ab202554 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval 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-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Adenine Nucleotide Translocator 2/ANT 2 with ab325120 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human kidney.
The section was incubated with ab325120 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-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling Adenine Nucleotide Translocator 2/ANT 2 with ab325120 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : weak staining on human skeletal muscle.
The section was incubated with ab325120 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-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Adenine Nucleotide Translocator 2/ANT 2 with ab325120 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human liver.
The section was incubated with ab325120 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-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Adenine Nucleotide Translocator 2/ANT 2 with ab325120 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse liver (PMID : 27048932, PMID : 31489369).
The section was incubated with ab325120 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-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Adenine Nucleotide Translocator 2/ANT 2 with ab325120 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse kidney (PMID : 27048932, PMID : 31489369).
The section was incubated with ab325120 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-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling Adenine Nucleotide Translocator 2/ANT 2 with ab325120 at 1/100 (5.04 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : weak staining on mouse skeletal muscle (PMID : 27048932, PMID : 31489369).
The section was incubated with ab325120 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

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse kidney tissue staining Adenine Nucleotide Translocator 2/ANT 2 with ab325120 at a 1 : 100 (5.04 ug/ml) dilution, ab202554 anti-COX IV used at 1 : 500 (0.23 ug/ml) dilution.

Panel A : merged staining of anti-ANT 2 (green; Opal™520) and anti-COX IV (magenta; Opal™570) on mouse kidney.
Panel B : anti-ANT 2 showed mitochondrial staining in mouse kidney.
Panel C : anti-COX IV showed mitochondrial staining in mouse 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 ab325120 and ab202554 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

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

• WB

Supplier Data

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

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

Lane 1 is incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 and lanes 2-4 are incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

Exposure time : Lane1 : 169 seconds, lanes2-4 : 10 seconds

All lanes:

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (ab325120) at 1/1000 dilution

Lane 1:

Rat kidney tissue lysate at 40 µg

Lane 2:

Human colon tissue lysate at 40 µg

Lane 3:

Human lung tissue lysate at 40 µg

Lane 4:

Human placenta tissue lysate at 40 µg

Secondary

Lane 1:

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

Lanes 2 - 4:

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

Observed band size: 32 kDa

false

• WB

Supplier Data

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

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

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

All lanes:

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (ab325120) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole lysate at 20 µg

Lane 2:

HeLa non-mitochondrial fraction at 20 µg

Lane 3:

HeLa mitochondrial fraction at 20 µg

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 32 kDa,36 kDa,16 kDa

false

Exposure time: 125s

• WB

Supplier Data

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

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

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

All lanes:

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (ab325120) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting SLC25A5 whole cell lysate at 20 µg

Lane 3:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 5:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 6:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 32 kDa,36 kDa

false

Exposure time: 125s

• WB

Supplier Data

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

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

Low expression : skeletal muscle(PMID : 27048932)

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

All lanes:

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (ab325120) at 1/1000 dilution

Lane 1:

Mouse colon tissue lysate at 40 µg

Lane 2:

Mouse kidney tissue lysate at 40 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 40 µg

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 32 kDa,36 kDa

false

Exposure time: 169s

• WB

Supplier Data

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (AB325120)

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

293T cells express endogenously SLC25A5. This antibody does not cross-react with human SLC25A4 by WB.

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

All lanes:

Western blot - Anti-Adenine Nucleotide Translocator 2/ANT 2 antibody [EPR29813-534] (ab325120) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 2:

293T transfected with human SLC25A5 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T transfected with human SLC25A4 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 32 kDa,34 kDa,36 kDa

false

Exposure time: 125s