Anti-ENT1 antibody [EPR30527-554]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENT1 antibody [EPR30527-554] (AB324975)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling ENT1 with ab324975 at 1/1000 (0.52 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse spleen.
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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENT1 antibody [EPR30527-554] (AB324975)

Immunohistochemical analysis of paraffin-embedded Mouse breast cancer tissue labeling ENT1 with ab324975 at 1/1000 (0.52 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse breast cacner.
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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENT1 antibody [EPR30527-554] (AB324975)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling ENT1 with ab324975 at 1/1000 (0.52 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining 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

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Immunocytochemistry/ Immunofluorescence - Anti-ENT1 antibody [EPR30527-554] (AB324975)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized MEF (mouse embryo fibroblast) cells labelling ENT1 with ab324975 at 1/200 (2.6 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 membranous and cytoplasmic staining in MEF cell line (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).

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

-ve control 1 : ab324975 at 1/200 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• WB

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Western blot - Anti-ENT1 antibody [EPR30527-554] (AB324975)

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-ENT1 antibody [EPR30527-554] (ab324975) at 1/1000 dilution

Lane 1:

MEF (mouse embryo fibroblast) transfected with scrambled siRNA control whole cell lysate at 5 µg

Lane 2:

MEF transfected with siRNA specifically targeting SLC29A1 whole cell lysate at 5 µg

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 50 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-ENT1 antibody [EPR30527-554] (AB324975)

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

Low expression : skeletal muscle(PMID : 11027664)

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

All lanes:

Western blot - Anti-ENT1 antibody [EPR30527-554] (ab324975) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 40 µg

Lane 2:

Mouse skeletal muscle tissue lysate at 40 µg

Lane 3:

Mouse lung tissue lysate at 40 µg

Secondary

All lanes:

Western blot at 1/20000 dilution

Observed band size: 50 kDa,36 kDa

false

Exposure time: 81s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENT1 antibody [EPR30527-554] (AB324975)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling ENT1 with ab324975 at 1/1000 (0.52 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining in mouse skeletal muscle.
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