Anti-EAAT2 antibody [EPR19794] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT2 antibody [EPR19794] - BSA and Azide free (AB251423)

This data was developed using ab205247, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling EAAT2 with ab205247 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on neurons of mouse liver is observed [PMID : 19718708].

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT2 antibody [EPR19794] - BSA and Azide free (AB251423)

This data was developed using ab205247, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling EAAT2 with ab205247 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative staining on mouse kidney.

Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT2 antibody [EPR19794] - BSA and Azide free (AB251423)

This data was developed using ab205247, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labeling EAAT2 with ab205247 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on neurons of mouse hippocampus is observed [PMID : 25391854].

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-EAAT2 antibody [EPR19794] - BSA and Azide free (AB251423)

This data was developed using ab205247, the same antibody clone in a different buffer formulation.

EAAT2 was immunoprecipitated from 0.35 mg of Rat brain lysate with ab205247 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab205247 at 1/2000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Rat brain lysate, 10μg (Input).
Lane 2 : ab205247 IP in Rat brain lysate.
Lane 3 : Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab205247 in Rat brain lysate.

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

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-EAAT2 antibody [EPR19794] (<a href='/en-us/products/primary-antibodies/eaat2-antibody-epr19794-ab205247'>ab205247</a>)

Predicted band size: 62 kDa

false

• WB

Supplier Data

Western blot - Anti-EAAT2 antibody [EPR19794] - BSA and Azide free (AB251423)

This data was developed using ab205247, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

EAAT2 is a multi-pass membrane protein, the main transporter that clears the excitatory neurotransmitter glutamate in the central nervous system. The bands around 180KD are multimers of EAAT2, which is consistent with what have been described in the literatures (PMID : 24569372 and 20193040).

All lanes:

Western blot - Anti-EAAT2 antibody [EPR19794] (<a href='/en-us/products/primary-antibodies/eaat2-antibody-epr19794-ab205247'>ab205247</a>) at 1/2000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Rat brain tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Mouse spleen tissue lysate at 20 µg

Lane 5:

Rat liver tissue lysate at 20 µg

Lane 6:

Rat kidney tissue lysate at 20 µg

Lane 7:

Rat spleen tissue lysate at 20 µg

Lane 8:

C6 (Rat glial tumor cell line) whole cell lysate at 20 µg

Lane 9:

Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate at 20 µg

Lane 10:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg

Lane 11:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 20 µg

Lane 12:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell 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 1/100000 dilution

Predicted band size: 62 kDa

Observed band size: 180 kDa,65 kDa

false

Exposure time: 5s