Anti-SLC7A5/LAT1 antibody [EPR29892-591] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-SLC7A5/LAT1 antibody [EPR29892-591] - BSA and Azide free (AB325953)

This data was developed using ab324354, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse hippocampus tissue staining SLC7A5/LAT1 with ab324354 at a 1/2000 dilution ab236870 anti-NeuN/RBFOX3 used at 1/10000 dilution and ab254264 anti-MAP2 used at a 1/4000 dilution.

Panel A : anti-SLC7A5/LAT1 (green; Opal™520) anti-NeuN/RBFOX3 (magenta; Opal™690) and anti-MAP2 (gray; Opal™570) on mouse cerebrum.

Panel B : anti-SLC7A5/LAT1 staining blood vessels in mouse cerebrum.

Panel C : anti-NeuN/RBFOX3 staining nucleus of neurons in mouse cerebrum.

Panel D : anti-MAP2 staining cell body and dendrites of neurons in mouse cerebrum.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324354 ab236870 and ab254264 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.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-SLC7A5/LAT1 antibody [EPR29892-591] - BSA and Azide free (AB325953)

This data was developed using ab324354, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse hippocampus tissue staining SLC7A5/LAT1 with ab324354 at a 1/2000 dilution ab236870 anti-NeuN/RBFOX3 used at 1/10000 dilution and ab254264 anti-MAP2 used at a 1/4000 dilution.

Panel A : anti-SLC7A5/LAT1 (green; Opal™520) anti-NeuN/RBFOX3 (magenta; Opal™690) and anti-MAP2 (gray; Opal™570) on mouse hippocampus.

Panel B : anti-SLC7A5/LAT1 staining blood vessels in mouse hippocampus.

Panel C : anti-NeuN/RBFOX3 staining nucleus of neurons in mouse hippocampus.

Panel D : anti-MAP2 staining cell body and dendrites of neurons in mouse hippocampus.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324354 ab236870 and ab254264 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.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-SLC7A5/LAT1 antibody [EPR29892-591] - BSA and Azide free (AB325953)

This data was developed using ab324354, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse pancreas tissue staining SLC7A5/LAT1 with ab324354 at a 1/2000 dilution ab278044 anti-Carboxypeptidase A used at 1/4000 dilution and ab181547 anti-Insulin used at a 1/20000 dilution.

Panel A : anti-SLC7A5/LAT1 (green; Opal™520) anti-Carboxypeptidase A (magenta; Opal™690) and anti-Insulin (gray; Opal™570) on mouse pancreas.

Panel B : anti-SLC7A5/LAT1 staining membrane in mouse pancreas.

Panel C : anti-Carboxypeptidase A staining acinar cells in mouse pancreas.

Panel D : anti-Insulin staining islet cells in mouse pancreas.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324354 ab278044 and ab181547 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.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-SLC7A5/LAT1 antibody [EPR29892-591] - BSA and Azide free (AB325953)

This data was developed using ab324354, the same antibody clone in a different buffer formulation.

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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC7A5/LAT1 antibody [EPR29892-591] - BSA and Azide free (AB325953)

This data was developed using ab324354, the same antibody clone in a different buffer formulation.

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC7A5/LAT1 antibody [EPR29892-591] - BSA and Azide free (AB325953)

This data was developed using ab324354, the same antibody clone in a different buffer formulation.

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC7A5/LAT1 antibody [EPR29892-591] - BSA and Azide free (AB325953)

This data was developed using ab324354, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (AB325953)

This data was developed using ab324354, the same antibody clone in a different buffer formulation.

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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SLC7A5/LAT1 antibody [EPR29892-591] - BSA and Azide free (AB325953)

This data was developed using ab324354, the same antibody clone in a different buffer formulation.

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.

• WB

Supplier Data

Western blot - Anti-SLC7A5/LAT1 antibody [EPR29892-591] - BSA and Azide free (AB325953)

This data was developed using ab324354, the same antibody clone in a different buffer formulation.

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] (<a href='/en-us/products/primary-antibodies/slc7a5-lat1-antibody-epr29892-591-ab324354'>ab324354</a>) 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] - BSA and Azide free (AB325953)

This data was developed using ab324354, the same antibody clone in a different buffer formulation.

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] (<a href='/en-us/products/primary-antibodies/slc7a5-lat1-antibody-epr29892-591-ab324354'>ab324354</a>) 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] - BSA and Azide free (AB325953)

This data was developed using ab324354, the same antibody clone in a different buffer formulation.

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] (<a href='/en-us/products/primary-antibodies/slc7a5-lat1-antibody-epr29892-591-ab324354'>ab324354</a>) 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

• WB

Supplier Data

Western blot - Anti-SLC7A5/LAT1 antibody [EPR29892-591] - BSA and Azide free (AB325953)

This data was developed using ab324354, 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.

All lanes:

Western blot - Anti-SLC7A5/LAT1 antibody [EPR29892-591] (<a href='/en-us/products/primary-antibodies/slc7a5-lat1-antibody-epr29892-591-ab324354'>ab324354</a>) 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