Anti-EAAT2 antibody [EPR19798]

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-EAAT2 antibody [EPR19798] (AB205248)

Immunohistochemical analysis of 4% paraformaldehyde-fixed 0.2% Triton X-100 permeabilized frozen Mouse striatum tissue labeling EAAT2 with ab205248 at 1/500 dilution followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Membrane staining on mouse striatum is observed [PMID : 25391854]. The nuclear counterstain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is ab150077 at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT2 antibody [EPR19798] (AB205248)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling EAAT2 with ab205248 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative control : no staining on mouse kidney [PMID : 11038258].

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 [EPR19798] (AB205248)

Immunohistochemical analysis of paraffin-embedded mouse striatum tissue labeling EAAT2 with ab205248 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane staining on mouse striatum 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EAAT2 antibody [EPR19798] (AB205248)

Immunohistochemical analysis of paraffin-embedded rat striatum tissue labeling EAAT2 with ab205248 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane staining on Rat striatum is observed. 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 [EPR19798] (AB205248)

EAAT2 was immunoprecipitated from 0.35 mg of Mouse brain lysate with ab205248 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab205248 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Mouse brain lysate, 10μg (Input).

Lane 2 : ab205248 IP in Mouse brain lysate.

Lane 3 : Rabbit IgG,monoclonal [EPR25A]- Isotype Control (ab172730) instead of ab205248 in Mouse brain lysate.

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

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-EAAT2 antibody [EPR19798] (ab205248)

Predicted band size: 62 kDa

false

• WB

Supplier Data

Western blot - Anti-EAAT2 antibody [EPR19798] (AB205248)

Blocking/Dilution buffer : 5% NFDM/TBST.

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

2. EATT2 is primarily expressed in astrocytes but is also expressed in neurons of the retina and during fetal development(PMID : 12176072).

All lanes:

Western blot - Anti-EAAT2 antibody [EPR19798] (ab205248) at 1/2000 dilution

Lane 1:

Mouse brain lysate at 20 µg

Lane 2:

Mouse liver lysate at 20 µg

Lane 3:

Mouse kidney lysate at 20 µg

Lane 4:

Rat brain lysate at 20 µg

Lane 5:

Rat liver lysate at 20 µg

Lane 6:

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

Lane 7:

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

Lane 8:

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

Lane 9:

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

Lane 10:

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: 1s