Anti-NeuN antibody [EPR12763] - Neuronal Marker - Rat IgG2a (Chimeric)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker - Rat IgG2a (Chimeric) (AB279297)

IHC image of NeuN staining in a section of formalin-fixed paraffin-embedded normal human cerebral cortex performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab279297, 1ug/ml, for 15 mins at room temperature. A rabbit anti-rat IgG2a, ab102248, was added for 8 mins at room temperature and detected using an HRP conjugated goat anti-rabbit compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC

Unknown

Immunocytochemistry - Anti-NeuN antibody [EPR12763] - Neuronal Marker - Rat IgG2a (Chimeric) (AB279297)

Immunofluorescence staining of NeuN using ab279297 in human SHSY5Y cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab279297 at 1.0 μg/ml. Cells were then incubated with ab150165, Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and nuclear DNA was labelled with DAPI (shown in blue). The secondary only control (bottom row) was not incubated with ab279297 but otherwise processed the same. Images were acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC

Unknown

Immunocytochemistry - Anti-NeuN antibody [EPR12763] - Neuronal Marker - Rat IgG2a (Chimeric) (AB279297)

Immunofluorescence staining of NeuN using ab279297 in human SHSY5Y cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab279297 at 1.0 μg/ml. Cells were then incubated with ab150165, Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and nuclear DNA was labelled with DAPI (shown in blue). The secondary only control (bottom row) was not incubated with ab279297 but otherwise processed the same. Images were acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-NeuN antibody [EPR12763] - Neuronal Marker - Rat IgG2a (Chimeric) (AB279297)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized rat primary neural/glia cells labelling NeuN with ab279297 at 1/1000 dilution (0.1μg)/ Right compared with a Mouse monoclonal IgG isotype control/ Left.

Goat F(ab)2 Anti-Rat IgG Fc (Alexa Fluor^® 647, ab150163) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NeuN antibody [EPR12763] - Neuronal Marker - Rat IgG2a (Chimeric) (AB279297)

Composite multiplex immunofluorescence staining of ab279297, ab317042 and ab308439 staining NeuN, NEFH and Tau (MBD region) in Mouse Primary Neurons DIV14 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab279297 (shown in green), ab317042 (shown in magenta) and ab308439 (shown in yellow) at 1µg/ml. Cells were then incubated with ab150161 Goat F(ab')2 Anti-Rat IgG Fc (Alexa Fluor® 488) preadsorbed, ab150175 Goat Anti-Chicken IgY H&L (Alexa Fluor® 647) preadsorbed and ab150084 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• WB

Supplier Data

Western blot - Anti-NeuN antibody [EPR12763] - Neuronal Marker - Rat IgG2a (Chimeric) (AB279297)

Exposure time : Lane 1 : 48 seconds; Lane 2, 3 : 4.5 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-NeuN antibody [EPR12763] - Neuronal Marker - Rat IgG2a (Chimeric) (ab279297) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Rat brain tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 34 kDa

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