Anti-SLC7A5/LAT1 antibody [EPR29892-591]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SLC7A5/LAT1 antibody [EPR29892-591] (AB324354)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labelling SLC7A5/LAT1 with ab324354 at 1/500 (1.024 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 membranous staining in F9 cell line and weak staining in mIMCD3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : mIMCD3.
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 (1 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 (2 ug/ml) dilution (Magenta).

-ve control 1 : ab324354 at 1/2000 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution. -ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC7A5/LAT1 antibody [EPR29892-591] (AB324354)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling SLC7A5/LAT1 with ab324354 at 1/2000 (0.256 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on blood vessels of mouse cerebrum (PMID : 11011012). 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.

• Flow Cyt

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Flow Cytometry - Anti-SLC7A5/LAT1 antibody [EPR29892-591] (AB324354)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized mIMCD3 (mouse inner medullary collecting duct epithelial cell, Left) / F9 (mouse embryonal carcinoma epithelial cell, Right) cells labelling SLC7A5/LAT1 with ab324354 at 1/5000 dilution (0.01ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : mIMCD3.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC7A5/LAT1 antibody [EPR29892-591] (AB324354)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling SLC7A5/LAT1 with ab324354 at 1/2000 (0.256 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse pancreas. 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-SLC7A5/LAT1 antibody [EPR29892-591] (AB324354)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling SLC7A5/LAT1 with ab324354 at 1/2000 (0.256 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse testis. 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.

• WB

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Western blot - Anti-SLC7A5/LAT1 antibody [EPR29892-591] (AB324354)

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

Low expression : brown fat.

The identity of the band higher than 50 kDa is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-SLC7A5/LAT1 antibody [EPR29892-591] (ab324354) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 50 µg

Lane 2:

Mouse brown fat lysate at 50 µg

Secondary

All lanes:

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

Observed band size: 37 kDa,124 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-SLC7A5/LAT1 antibody [EPR29892-591] (AB324354)

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

Low expression : liver (PMID : 27912058).

The identity of the band higher than 50 kDa is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-SLC7A5/LAT1 antibody [EPR29892-591] (ab324354) at 1/1000 dilution

Lane 1:

Mouse ovary tissue lysate at 72 µg

Lane 2:

Mouse liver tissue lysate at 72 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: 37 kDa,124 kDa

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-SLC7A5/LAT1 antibody [EPR29892-591] (AB324354)

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-SLC7A5/LAT1 antibody [EPR29892-591] (ab324354) at 1/1000 dilution

Lane 1:

F9 (mouse embryonal carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 40 µg

Lane 2:

F9 transfected with siRNA specifically targeting SLC7A5/LAT1 whole cell 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: 37 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-SLC7A5/LAT1 antibody [EPR29892-591] (AB324354)

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

Low expression : RAW 264.7, mIMCD3.

The identity of the bands higher than 50 kDa are unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-SLC7A5/LAT1 antibody [EPR29892-591] (ab324354) at 1/1000 dilution

Lane 1:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 72 µg

Lane 2:

P815 (mouse mastocytoma mast cell) whole cell lysate at 72 µg

Lane 3:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 72 µg

Lane 4:

mIMCD3 (mouse inner medullary collecting duct epithelial cell) whole cell lysate at 72 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: 37 kDa,124 kDa

false

Exposure time: 6s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC7A5/LAT1 antibody [EPR29892-591] (AB324354)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling SLC7A5/LAT1 with ab324354 at 1/2000 (0.256 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on 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.