Anti-EAAT3 antibody [EPR25149-62]

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat kidney (fresh) tissue labeling EAAT3 with ab288441 at 1/500 (0.982 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Positive staining on the proximal convoluted tubule of rat kidney is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat hippocampus (fresh) tissue labeling EAAT3 with ab288441 at 1/50 (9.82 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Positive staining on rat hippocampus is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling EAAT3 with ab288441 at 1/1000 (0.491 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Cytoplasmic staining on rat cerebrum (PMID : 11242046). The section was incubated with ab288441 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 Leica DS9800 (BOND™, Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labelling EAAT3 with ab288441 at 1/1000 (0.491 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Apical staining on proximal convoluted tubules in mouse kidney (PMID : 9435692).The section was incubated with ab288441 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 Leica DS9800 (BOND™, Polymer Refine Detection) was used.

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling EAAT3 with ab288441 at 1/100 (4.91 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in mouse primary neuron cells. Confocal scanning Z step was set as 0.3 um followed by image processing with maximum Z projection. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain neuron-specific cytoskeletal protein at 1/500 (4ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2ug/ml) dilution.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver (fresh) (-) tissue labeling EAAT3 with ab288441 at 1/50 (9.82 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). No staining on rat liver (Negative control : PMID 11242046) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse kidney (fresh) tissue labeling EAAT3 with ab288441 at 1/500 (0.982 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Positive staining on the proximal convoluted tubule of mouse kidney is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labelling EAAT3 with ab288441 at 1/1000 (0.491 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Negative control : no staining on rat testis.The section was incubated with ab288441 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 Leica DS9800 (BOND™, Polymer Refine Detection) was used.

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-EAAT3 antibody [EPR25149-62] (AB288441)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labelling EAAT3 with ab288441 at 1/1000 (0.491 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Apical staining on proximal convoluted tubules in rat kidney (PMID : 9435692).The section was incubated with ab288441 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 Leica DS9800 (BOND™, Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (fresh) (-) tissue labeling EAAT3 with ab288441 at 1/50 (9.82 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). No staining on mouse liver (Negative control : PMID 11242046) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labelling EAAT3 with ab288441 at 1/1000 (0.491 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Cytoplasmic staining on mouse hippocampus (PMID : 11242046).The section was incubated with ab288441 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 Leica DS9800 (BOND™, Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling EAAT3 with ab288441 at 1/1000 (0.491 ug/ml) followed by a ready to use Leica DS9800 (BOND™, Polymer Refine Detection) was used. Negative control : no staining on mouse spleen.The section was incubated with ab288441 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 Leica DS9800 (BOND™, Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebellum (fresh) tissue labeling EAAT3 with ab288441 at 1/50 (9.82 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Positive staining on mouse cerebellum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IP

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Immunoprecipitation - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

EAAT3 was immunoprecipitated from 0.35 mg Rat hippocampus tissue lysate 10 ug with ab288441 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288441 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Rat hippocampus tissue lysate 10 ug

Lane 2 : ab288441 IP in Rat hippocampus tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288441 in rat hippocampus tissue lysate

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

Exposure time : 8 seconds

All lanes:

Immunoprecipitation - Anti-EAAT3 antibody [EPR25149-62] (ab288441)

Predicted band size: 57 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

EAAT3 was immunoprecipitated from 0.35 mg Mouse hippocampus tissue lysate 10 ug with ab288441 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288441 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse hippocampus tissue lysate 10 ug

Lane 2 : ab288441 IP in Mouse hippocampus tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288441 in mouse hippocampus tissue lysate

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

Exposure time : 8 seconds

All lanes:

Immunoprecipitation - Anti-EAAT3 antibody [EPR25149-62] (ab288441)

Predicted band size: 57 kDa

false

• WB

Lab

Western blot - Anti-EAAT3 antibody [EPR25149-62] (AB288441)

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

Exposure time : Lanes 1-4 : 26 seconds; Lanes 5-7 : 3 minutes

Monomers : 50-75 kda; Aggregates : 150-250 kda
The MW and expression pattern is consistent to the literature (PMID : 30840898)

All lanes:

Western blot - Anti-EAAT3 antibody [EPR25149-62] (ab288441) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Lane 4:

Mouse testis tissue lysate at 20 µg

Lane 5:

Rat hippocampus tissue lysate at 20 µg

Lane 6:

Rat brain tissue lysate at 20 µg

Lane 7:

Rat testis tissue 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

Predicted band size: 57 kDa

Observed band size: 150-250 kDa,50-75 kDa

false