All lanes : Anti-Glutamate Receptor 1 (AMPA subtype) antibody [EPR19522] (ab183797) at 1/5000 dilution

Lane 1 : Mouse Glutamate Receptor 1 fragment recombinant protein
Lane 2 : Mouse Glutamate Receptor 2 fragment recombinant protein

Lysates/proteins at 0.01 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 102 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure times: Lane 1: 2 seconds; Lane 2: 3 minutes.

Mouse Glutamate Receptor 1 fragment recombinant protein contains aa19-184 with a GST/His-Tag^®. Mouse Glutamate Receptor 2 fragment recombinant protein contains aa25-288 with a His-Tag^®.

All lanes : Anti-Glutamate Receptor 1 (AMPA subtype) antibody [EPR19522] (ab183797) at 1/1000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Rat liver lysate
Lane 3 : Mouse hippocampus lysate
Lane 4 : Mouse cerebellum lysate
Lane 5 : Rat brain lysate
Lane 6 : Rat cerebellum lysate
Lane 7 : Rat hippocampus lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 102 kDa
Observed band size: 102 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure times: Lane 1 and 2: 30 seconds; Lane 3, 4, 5, 6 and 7: 15 seconds.

Rodent Glutamate Receptor 1 is widely expressed in brain and represents the predominant excitatory neurotransmitter system but not in liver.

Negative control: Rat liver (PMID: 2480522).

All lanes : Anti-Glutamate Receptor 1 (AMPA subtype) antibody [EPR19522] (ab183797) at 1/1000 dilution

Lane 1 : Human cerebellum lysate
Lane 2 : Human muscle lysate
Lane 3 : Human fetal brain lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 102 kDa
Observed band size: 102 kDa


Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

Human Glutamate Receptor 1 is widely expressed in brain and represents the predominant excitatory neurotransmitter system but not in muscle.

Negative control: human muscle (PMID: 1652753).

Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labeling Glutamate Receptor 1 (AMPA subtype) with ab183797 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on mouse hippocampus was observed [PMID: 15723058]. 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.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Glutamate Receptor 1 (AMPA subtype) with ab183797 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. No staining on mouse liver is observed. Counter stained with Hematoxylin.

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

Immunohistochemical analysis of paraffin-embedded Rat hippocampus tissue labeling Glutamate Receptor 1 (AMPA subtype) with ab183797 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on rat hippocampus is observed [PMID: 15723058]. 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.

Immunohistochemical analysis of paraffin-embedded Rat stomach tissue labeling Glutamate Receptor 1 (AMPA subtype) with ab183797 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. No staining on rat stomach is observed. Counter stained with Hematoxylin.

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

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus tissue labeling Glutamate Receptor 1 (AMPA subtype) with ab183797 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasm staining on mouse hippocampus was observed.  The nuclear counterstain is DAPI (blue).

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

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus tissue labeling Glutamate Receptor 1 (AMPA subtype) with ab183797 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasm staining on mouse hippocampus was observed.  The nuclear counterstain is DAPI (blue).

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

Glutamate Receptor 1 (AMPA subtype) was immunoprecipitated from 0.35 mg of Mouse brain whole cell lysate with ab183797 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab183797 at 1/1000 dilution. VeriBlot for IP Detection Reaction (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: Mouse brain whole cell lysate, 10ug (Input).

Lane 2: ab183797 IP in Mouse brain whole cell lysate.

Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab183797 in Mouse brain whole cell lysate.

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

Exposure time: 3 minutes.