Anti-NeuN antibody [EPR21906] - Neuronal Marker

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR21906] - Neuronal Marker (AB236870)

Immunohistochemical analysis of paraffin-embedded human cerebellum tissue labeling NeuN with ab236870 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining in granule cells of human cerebellum (PMID : 8813082; PMID : 1483388). Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Heat mediated antigen retrieval was performed 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-NeuN antibody [EPR21906] - Neuronal Marker (AB236870)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling NeuN with ab236870 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining in neurons of human cerebrum (PMID : 8813082; PMID : 1483388; PMID : 26085943). Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-NeuN antibody [EPR21906] - Neuronal Marker (AB236870)

NeuN was immunoprecipitated from 0.35 mg human cerebellum lysate with ab236870 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab236870 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : Human cerebellum lysate 10 μg (Input).
Lane 2 : ab236870 IP in human cerebellum lysate (+).
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab236870 in human cerebellum lysate (-).

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 30 seconds.

The molecular weight observed is consistent with what has been described in the literature (PMID : 1483388; PMID : 26085943).

All lanes:

Immunoprecipitation - Anti-NeuN antibody [EPR21906] - Neuronal Marker (ab236870)

Predicted band size: 34 kDa

false

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-NeuN antibody [EPR21906] - Neuronal Marker (AB236870)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebellum tissue labeling NeuN with ab236870 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasmic and nuclear staining in rat cerebellum (PMID : 19713214).

The nuclear counter stain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-NeuN antibody [EPR21906] - Neuronal Marker (AB236870)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse hippocampus tissue staining SLC7A5/LAT1 with ab324354 at a 1/2000 dilution, ab236870 anti-NeuN/RBFOX3 used at 1/10000 dilution and ab254264 anti-MAP2 used at a 1/4000 dilution.

Panel A : anti-SLC7A5/LAT1 (green; Opal™520), anti-NeuN/RBFOX3 (magenta; Opal™690) and anti-MAP2 (gray; Opal™570) on mouse hippocampus.

Panel B : anti-SLC7A5/LAT1 staining blood vessels in mouse hippocampus.

Panel C : anti-NeuN/RBFOX3 staining nucleus of neurons in mouse hippocampus.

Panel D : anti-MAP2 staining cell body and dendrites of neurons in mouse hippocampus.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324354, ab236870 and ab254264 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR21906] - Neuronal Marker (AB236870)

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling NeuN with ab236870 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining in neurons of rat cerebrum (PMID : 8813082; PMID : 1483388; PMID : 26085943). Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-NeuN antibody [EPR21906] - Neuronal Marker (AB236870)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse hippocampus tissue staining SLC7A5/LAT1 with ab324354 at a 1/2000 dilution, ab236870 anti-NeuN/RBFOX3 used at 1/10000 dilution and ab254264 anti-MAP2 used at a 1/4000 dilution.

Panel A : anti-SLC7A5/LAT1 (green; Opal™520), anti-NeuN/RBFOX3 (magenta; Opal™690) and anti-MAP2 (gray; Opal™570) on mouse cerebrum.

Panel B : anti-SLC7A5/LAT1 staining blood vessels in mouse cerebrum.

Panel C : anti-NeuN/RBFOX3 staining nucleus of neurons in mouse cerebrum.

Panel D : anti-MAP2 staining cell body and dendrites of neurons in mouse cerebrum.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324354, ab236870 and ab254264 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR21906] - Neuronal Marker (AB236870)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling NeuN with ab236870 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear and cytoplasmic staining in neurons of mouse cerebrum (PMID : 8813082; PMID : 1483388; PMID : 26085943). Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-NeuN antibody [EPR21906] - Neuronal Marker (AB236870)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebellum tissue labeling NeuN with ab236870 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasmic and nuclear staining in mouse cerebellum (PMID : 19713214).

The nuclear counter stain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• WB

Lab

Western blot - Anti-NeuN antibody [EPR21906] - Neuronal Marker (AB236870)

Western blot : Rabbit Monoclonal[EPR21906] to NeuN ab236870 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 40 kDa in Wild-type A549 cell lysates with no signal observed at this size in RBFOX3 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-NeuN antibody [EPR21906] - Neuronal Marker (ab236870) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

RBFOX3 knockout A549 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 34 kDa

Observed band size: 40 kDa,42 kDa

false

• WB

Supplier Data

Western blot - Anti-NeuN antibody [EPR21906] - Neuronal Marker (AB236870)

Exposure times : Lanes 1-2 : 3 minutes; Lane 3 : 92 seconds; Lanes 4-7 : 26 seconds; Lanes 8-11 : 15 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 1483388; PMID : 26085943).

All lanes:

Western blot - Anti-NeuN antibody [EPR21906] - Neuronal Marker (ab236870) at 1/1000 dilution

Lane 1:

Human fetal brain lysate at 20 µg

Lane 2:

Human hippocampus lysate at 20 µg

Lane 3:

Human cerebellum lysate at 20 µg

Lane 4:

Mouse brain lysate at 20 µg

Lane 5:

Mouse cerebral cortex lysate at 20 µg

Lane 6:

Mouse cerebellum lysate at 20 µg

Lane 7:

Mouse hippocampus lysate at 20 µg

Lane 8:

Rat brain lysate at 20 µg

Lane 9:

Rat cerebral cortex lysate at 20 µg

Lane 10:

Rat cerebellum lysate at 20 µg

Lane 11:

Rat hippocampus 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: 34 kDa

Observed band size: 46 kDa,48 kDa

false