Anti-nNOS (neuronal) antibody [EP1855Y] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-nNOS (neuronal) antibody [EP1855Y] - BSA and Azide free (AB271857)

Overlay histogram showing PC-12 cells stained with unpurified ab76067 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (unpurified ab76067, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in PC-12 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.Please note that Abcam do not have any data for use of this antibody in non-fixed cells. We welcome any customer feedback.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-nNOS (neuronal) antibody [EP1855Y] - BSA and Azide free (AB271857)

Intracellular Flow Cytometry analysis of PC-12 cells labelling nNos (neuronal) with unpurified ab76067 at 1/15 (red). Cells were fixed with 100% methanol. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-nNOS (neuronal) antibody [EP1855Y] - BSA and Azide free (AB271857)

Intracellular Flow Cytometry analysis of PC-12 cells labelling nNos (neuronal) with purified ab76067 at 1/600 (red). Cells were fixed with 100% methanol. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-nNOS (neuronal) antibody [EP1855Y] - BSA and Azide free (AB271857)

Immunocytochemistry analysis of PC-12 cells labelling nNOS (neuronal) (green) with purified ab76067 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counterstained with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red). Nuclear DNA was labelled with DAPI (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

• IP

Unknown

Immunoprecipitation - Anti-nNOS (neuronal) antibody [EP1855Y] - BSA and Azide free (AB271857)

ab76067 (unpurified) at 1/4 immunoprecipitating nNOS (neuronal) in rat brain tissue lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

All lanes:

Immunoprecipitation - Anti-nNOS (neuronal) antibody [EP1855Y] (<a href='/en-us/products/primary-antibodies/nnos-neuronal-antibody-ep1855y-ab76067'>ab76067</a>)

Predicted band size: 160 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-nNOS (neuronal) antibody [EP1855Y] - BSA and Azide free (AB271857)

ab76067 (purified) at 1/150 immunoprecipitating nNOS (neuronal) in rat brain tissue lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76067).

All lanes:

Immunoprecipitation - Anti-nNOS (neuronal) antibody [EP1855Y] (<a href='/en-us/products/primary-antibodies/nnos-neuronal-antibody-ep1855y-ab76067'>ab76067</a>)

Predicted band size: 160 kDa

false