Anti-NeuN antibody [EPR12763] - Neuronal Marker

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Immunohistochemical analysis of 4% PFA-fixed 0.2% Triton X-100 permeabilized frozen Mouse hippocampus (fresh frozen) tissue labeling MAP2 with ab318993 at 1/2000 (0.486 ug/ml) dilution followed by ab150169 Goat Anti-Chicken IgY H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/mL) dilution (Green).

Panel A : merged staining of anti-MAP2 (ab318993 green), anti-NeuN (ab177487 red) and anti-GFAP (ab313596 grey) on mouse hippocampus.
Panel B : anti-MAP2 stained on mouse hippocampus.
Panel C : anti-NeuN stained in neuron of mouse hippocampus.
Panel D : anti-GFAP stained in astrocytes of mouse hippocampus.
The nuclear counterstain was DAPI (Blue). The section was incubated with ab318993 ab177487 and ab313596 for 60 mins at room temperature. The section was then mounted using Fluoromount^®.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

Secondary antibody control : Secondary antibody is ab150169 Goat Anti-Chicken IgY H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/mL) dilution.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Fluorescence multiplex immunohistochemical analysis of human cerebellum tissue (formalin-fixed paraffin-embedded section).

Merged staining of Neu-N (ab177487; yellow; Opal™570), anti-beta III Tubulin (ab52623; red; Opal™690) and anti-GFAP (ab68428; green; Opal™520).

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ kit.

The section was incubated in three rounds of staining with ab177487 (1/1000 dilution), ab52623 (1/200 dilution) and ab68428 (1/250 dilution); each using a separate fluorescent tyramide signal amplification system.

Sodium citrate antigen retrieval (pH 6.0) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

DAPI (blue) was used as a nuclear counterstain.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Chromogenic multiplex immunohistochemical staining of FFPE normal human cerebellum tissue. ab177487, anti-NeuN DAB chromogen. ab68428, anti-GFAP purple chromogen and ab178846, anti- Iba1 teal chromogen plus haematoxylin counterstain.

Chromogenic immunostaining was performed on a Roche Ventana Discovery Ultra instrument. The section was deparaffinized and incubated with CC1 solution for 24 min 100°C. Following this with 3 rounds of staining in the order of ab177487 (1/600), ab178846 (1/4000) ab68428 (1/1000). Between rounds of staining, antibody denaturation was conducted using Ultra CC2 solution for 8 min at 100°C to avoid cross reactivity. Signal was developed with anti-rabbit HQ followed by anti-HQ HRP coupled with Chromomap DAB kit, Discovery purple or Discovery teal chromogens and haematoxylin II counterstain.

• ICC/IF

AbReview66918****

Immunocytochemistry/ Immunofluorescence - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Immunocytochemistry/immunofluorescence analysis of human neurons differentiated from iPSCs labelling NeuN (green) with ab177487 at 1/500 in 0.1% TritonX-100 1% goat serum 1X PBS for 16 hours at 4°C. Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100. Then cells were blocked with 5% serum for 20 minutes at 23°C. Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilutionwas used as the secondary antibody. Tuj1 antibody was used to stain neuronal dendrites and axons (red).

This image is courtesy of an Abreview submitted by Vladimir Milenkovic

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Immunohistochemical analysis of formalin fixed paraffin embedded human cerebellum labelling NeuN with ab177487 at a dilution of 1/500. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an ChromoMap DAB (RUO) IHC Detection Kit with anti rabbit HQ and anti HQ HRP. Heat mediated antigen retrieval was conducted for 24 min with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5. ab177487 was incubated at 37°C for 16 min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Immunofluorescence staining of NeuN using ab177487 in ioGlutamatergic Neurons (Human iPSC-Derived Glutamatergic Neurons, ab259259), which were differentiated for 1 day post induction.

The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Tween 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 ab177487 at 1 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Overlay histogram showing U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells stained with ab177487 (red line).

The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab177487 1/100 dilution) for 30 minutes at 22°C. The secondary antibody used was Alexa Fluor^®488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730 1μg/1x10⁶cells used under the same conditions. Unlabeled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

Alexa Fluor^® 488 (ab190195) and Alexa Fluor^® 647 (ab190565) conjugated versions are available for this clone.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gliocytoma tissue labelling NeuN with ab177487 at 1/3000 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

• IHC-P

AbReview38959****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

IHC-P image of FOX3/NeuN staining on sheep brain (Frontal cortex) sections using ab177487 (1/1000 dilution). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/1000 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250 dilution).

This image is courtesy of an abreview submitted by Carl Hobbs, Kings's College London, United Kingdom.

• IHC-P

AbReview38940****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

IHC-P image of FOX3/NeuN staining on mouse brain (frontal cortex) sections using ab177487 (1/800 dilution). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/800 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250 dilution).

This image is courtesy of an abreview submitted by Carl Hobbs, Kings's College London, United Kingdom.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Immunocytochemsitry/Immunofluorescence analysis of SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling NeuN (green) with ab177487 at 1/300 dilution. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/200 dilution) was used as the secondary antibody. Counterstained with DAPI (blue).

• IHC-P

AbReview38955****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

IHC-P image of FOX3/NeuN staining on zebrafish spinal cord sections using ab177487 (1/500). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/500 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250).

This image is courtesy of an abreview submitted by Carl Hobbs, Kings's College London, United Kingdom.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Immunocytochemistry/immunofluorescence analysis of Mouse primary neuron cells labelling NeuN with ab177487 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-MAP2 mouse monoclonal antibody (ab11267) at 1/200 dilution and visualised using Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) at 1/1000 dilution (red). Nuclear DNA was labelled with DAPI (blue).

Confocal image showing mainly nuclear staining in mouse primary neuron cells. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.

• IHC-P

AbReview38956****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

IHC-P image of FOX3/NeuN staining on cat cerebellum sections using ab177487 (1/1000 dilution).

Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/1000 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250 dilution).

This image is courtesy of an abreview submitted by Carl Hobbs, Kings's College London, United Kingdom.

• IHC (PFA fixed)

Supplier Data

Immunohistochemistry (PFA fixed) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

NeuN antibody ab177487 was used with Tissue Clearing Kit ab243298 to penetrate, stain and clear a 1 mm coronal section of mouse brain. Blue : DAPI, Green : NeuN. Learn more about tissue clearing kits, reagents, and protocols designed to make it easier to stain thick tissue sections and get more data from each valuable tissue section. For 1 mm brain sections, we recommend a starting dilution of 1 : 200, and also using Goat Anti-Rabbit IgG H&L AlexaFluor488 (ab150077) at a dilution of 1 : 400.

• IHC-P

AbReview38953****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

IHC-P image of FOX3/NeuN staining on rat brain (SVZ) sections using ab177487 (1/2000). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/2000 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250).

This image is courtesy of an abreview submitted by Carl Hobbs, Kings's College London, United Kingdom.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat hippocampus (fresh frozen) tissue labeling MAP2 with ab318993 at 1/2000 (0.486 ug/ml) dilution followed by ab150169 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) at 1/1000 2 ug/mL dilution (Green).

Panel A : merged staining of anti-MAP2 (ab318993, green), anti-NeuN (ab177487, red) and anti-GFAP (ab313596, grey) on rat hippocampus.
Panel B : anti-MAP2 stained on rat hippocampus.
Panel C : anti-NeuN stained in neuron of rat hippocampus.
Panel D : anti-GFAP stained in astrocytes of rat hippocampus.
The nuclear counterstain was DAPI (Blue). The section was incubated with ab318993, ab177487 and ab313596 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150169 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488)at 1/1000 2 ug/mL dilution.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

​An independent comparison of commercially available NeuN clones in IHC-Fr (acetone-fixed mouse dentate gyrus sections).

Competitor A : Leading mouse monoclonal.

Competitor B : Non-Abcam rabbit monoclonal.

ab177487 produces intense, specific staining with minimal background, even at half the dilution of competing antibodies.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat dorsal root ganglia staining GALR2 with ab271019 at a 1/2000 dilution and ab177487 anti-NeuN used at 1/300 dilution.

Panel A : merged staining of anti-GALR2 (green; Opal™520) and anti-NeuN (magenta; Opal™570) on Rat dorsal root ganglia.

Panel B : anti-GALR2 stained on cytoplasm of neurons.

Panel C : anti-NeuN stained on nucleus and cytoplasm of neurons.

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

The section was incubated in two rounds of staining : in the order of ab271019 and ab177487 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

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebrum tissue labelling NeuN with ab177487 at 1/100 dilution (B), SOX1 with ab242125 at 1/100 dilution (C) and Olig2 with ab109186 at 1/100 dilution (D). Anti-Rabbit and Mouse Polymer HRP was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. Heat mediated antigen retrieval (Leica ER2, PH9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibodies from the previous round, to avoid any cross-reactivity.

Panel A : merged staining of anti- NeuN (green, Opal™520), anti-SOX1 (red, Opal™570) and anti- Olig2 (yellow, Opal™690).

Panel B : anti-NeuN stained for neurons.

Panel C : anti-SOX1 stained on neural progenitors.

Panel D : anti-Olig2 stained on oligodendrocyte.

The section was incubated in three rounds of staining : in the order of ab177487, ab242125 and ab109186 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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Flow cytometry overlay histogram showing Neuro-2a cells stained with ab177487 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab177487) (1x 10⁶in 100μl at 5.0μg/ml (1/444)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody gave a positive signal in Neuro-2a Fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• IHC-Fr

AbReview42910****

Immunohistochemistry (Frozen sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

ab177487 staining NeuN in mouse brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with formaldehyde and blocked with Triton X-100 + 0.4% horse serum for 30 minutes at 20°C. Samples were incubated with primary antibody (1/500 in blocking solution) for 16 hours at 4°C. An Alexa Fluor^® 594-conjugated donkey anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

This image is courtesy of an Abreview submitted by Eva Borger

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

IHC-P image of NeuN (green) and GFAP (red) double staining on mouse cerebellum sections using ab177487 (1/5000) and ab4674 (1/1500) respectively.

The sections were deparaffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were then incubated with Rabbit Monoclonal to NeuN (ab177487) diluted at 1/5000 and Chicken Polyclonal to GFAP (ab4674) diluted at 1/1500. The primary antibody was detected using ab150097 Goat anti-rabbit IgG conjugated to Alexa Fluor^® 488 (1/500) and ab150176 Goat anti-chicken IgY conjugated to Alexa Fluor^® 594 (1/500)

This image is courtesy of an abreview submitted by Carl Hobbs, Kings's College London, United Kingdom.

• IHC-Fr

AbReview40650****

Immunohistochemistry (Frozen sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

ab177487 staining NeuN in mouse free floating 50 micron lumbar spinal cord tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with formaldehyde, permeabilized with Triton X-100 and blocked with 10% serum for 2 hours at 25°C. Samples were incubated with primary antibody (1/500 in PBS + Triton) for 16 hours at 4°C. An Alexa Fluor^® 594-conjugated donkey anti-rabbit IgG polyclonal (1/700) was used as the secondary antibody.

This image is courtesy of an Abreview submitted by Jianning Lu

• IHC-tissue clearing

Supplier Data

IHC-tissue clearing - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Anti-NeuN ab177487 was used with Tissue clearing kit – CUBIC (ab316246) and 3D Tissue Staining Kit – CUBIC (ab316248) to penetrate stain and clear a whole mouse brain.

White : nuclear staining
Red : NeuN.

Learn more about tissue clearing kits reagents and protocols designed to make it easier to stain whole brains and get more data from each valuable tissue sample. For a whole mouse brain we recommend starting with 10 ug of ab177487 and using a Fab fragment secondary antibody with 6.67 µg to create an antibody complex before 3D staining (see protocol for details). Additive A was used during the staining process.

The sample was imaged using a light-sheet microscope.

• WB

Lab

Western blot - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Exposure time -
Lane 1-2 : 3 minutes.
Lane 3-4 : 1 minute.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487) at 1/10000 dilution

Lane 1:

Human fetal brain tissue lysate at 10 µg

Lane 2:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 10 µg

Lane 3:

Mouse brain tissue lysate at 10 µg

Lane 4:

Rat brain tissue lysate at 10 µg

Secondary

All lanes:

Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 34 kDa

Observed band size: 46 kDa,48 kDa

false

• WB

Lab

Western blot - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

Different batches of ab177487 were tested on mouse brain lysate at 2.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 46,48 kDa.

All lanes:

Western blot - Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487)

Predicted band size: 34 kDa

false

• WB

Lab

Western blot - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

This blot was produced using 4-20% SDS-PAGE containing 15 μg of mouse brain lysate per lane at 150V for 1hr before being transferred onto a 0.45 μm PVDF membrane at 75V for 1hr. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab177487 (1/10,000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG (H+L) (ab97051) at 1/20,000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique.

ab177487 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected under these conditions. This data was generated as part of a structured antibody stability study supporting Abcam’s transition from cold‑chain to ambient shipping.

All lanes:

Western blot - Anti-NeuN antibody [EPR12763] - Neuronal Marker (ab177487) at 1/10000 dilution

All lanes:

Mouse brain lysate at 15 µg with NDFM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

false

Exposure time: 3s

• IHC-P

AbReview38954****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

IHC-P image of FOX3/NeuN staining on marmoset cerebellum sections using ab177487 (1/2000). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/2000 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250).

This image is courtesy of an abreview submitted by Carl Hobbs, Kings's College London, United Kingdom.

• IHC-P

AbReview38958****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

IHC-P image of FOX3/NeuN staining on goat cerebellum sections using ab177487 (1/500 dilution). Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/500 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250 dilution).

This image is courtesy of an abreview submitted by Carl Hobbs, Kings's College London, United Kingdom.

• IHC-P

AbReview38957****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

IHC-P image of FOX3/NeuN staining on dog cerebellum sections using ab177487 (1/500 dilution).

Sections were de-paraffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 minutes at 21°C. ab177487 was diluted 1/500 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to biotin (1/250 dilution).

This image is courtesy of an abreview submitted by Carl Hobbs, Kings's College London, United Kingdom.

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuN antibody [EPR12763] - Neuronal Marker (AB177487)

​An independent comparison of commercially available NeuN clones in IHC-P.

Competitor A : Leading mouse monoclonal.

Competitor B : Non-Abcam rabbit monoclonal.

Sodium citrate was used for antigen retrieval in all 3 samples.

ab177487 produces specific staining, equivalent to the leading mouse monoclonal at half the dilution. The non-Abcam mouse monoclonal was less specific as it stained Purkinje cells, which do not express NeuN.