Anti-beta III Tubulin antibody - Neuronal Marker

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

ab18207 staining beta III Tubulin in PC12 cells. The cells were fixed with 100% methanol (5 min) permeabilized with 0.1% PBS-Tween 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 ab18207 at 1µg/ml and ab7291 Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081 Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488) pre-adsorbed at 1/1000 dilution (shown in green) and ab150120 Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594) pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). 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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

Overlay histogram showing U-87MG (Human glioblastoma-astrocytoma epithelial cell line) cells stained with ab18207 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 20 min. 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 (ab18207 0.01μg/1x10⁶) for 30 min at 22°C. The secondary antibody used was Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab171870 0.01μg/1x10⁶ cells) used under the same conditions. Unlabelled 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.

This antibody gave a positive signal in U-87MG cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

Immunohistochemical analysis of formalin fixed paraffin embedded human cerebrum labelling beta III tubulin with ab18207 at a concentration of 0.32 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). ab18207 anti-beta III tubulin antibody was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

ab18207 staining beta III Tubulin in SK-N-SH (Human neuroblastoma cell line) cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab18207 at 1μg/ml and ab7291 at 1μg/ml overnight at +4°C followed by a further incubation at room temperature for 1h with a Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Negative controls : 1– Rabbit primary and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

ab18207 staining beta III Tubulin in Neuro-2a cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab18207 at 1μg/ml and ab7291 at 1μg/ml overnight at +4°C followed by a further incubation at room temperature for 1h with a Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Negative controls : 1– Rabbit primary and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

IHC image of ab18207 staining beta III Tubulin in rat cerebellum formalin fixed paraffin embedded tissue sections performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6 epitope retrieval solution 1) for 20 mins. The section was then incubated with ab18207 1 : 2000 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

Overlay histogram showing Neuro 2A cells stained with ab18207 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 20 min. 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 (ab18207 0.01μg/1x10⁶) for 30 min at 22°C. The secondary antibody used was Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081) at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab171870 0.01μg/1x10⁶ cells) used under the same conditions. Unlabelled 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.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

Immunohistochemical analysis of adult mice ovaries undergone Clarity processing staining tyrosine hydroxlase (TH) Beta III Tubulin (Tuj1) with ab18207 and brain derived neurotrpic factor (BDNF) with ab72439. Positive staing of Tuj1 and BDNF is evident in the theca cells and corpus luteum.

Courtesy of Feng Y et al. Sci Rep. 2017; 7: 44810. doi: 10.1038/srep44810 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

ab18207 at 1/2000 dilution staining mouse brain tissue sections by IHC-P. The tissue was formaldehyde fixed and an enzymatic antigen retrieval step was performed prior to incubation with the antibody for 16 hours. A biotinylated goat anti-rabbit IgG was used as the secondary.

This image is courtesy of Carl Hobbs, King's College London, United Kingdom

• ICC/IF

AbReview76541****

Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

Immunocytochemistry analysis of paraformaldehyde-fixed mouse brain staining with ab18207 at 1/200 dilution. Secondary antibody was Alexa Fluor® 568 Donkey Anti-Rabbit IgG. Samples were incubated with the primary antibody with 2% donkey serum in 0.2% TritonX 100 for 16 hours at 27°C. Blocking was done using 10% donkey serum for 2 hours at 28°C.

This image is courtesy of an Abreview submitted by Lenin Veeraval

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

ab18207 staining beta III Tubulin in primary rat neurons/glia DIV14 (prepared from E18 rat hippocampal brain area obtained from Transnetyx Tissue by BrainBits LLC cat.no. SDHEP) cells. The cells were fixed with 100% methanol (5 min) permeabilized with 0.1% PBS-Tween 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 ab18207 at 1µg/ml and ab7291 Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081 Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488) pre-adsorbed at 1/1000 dilution (shown in green) and ab150120 Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594) pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a high-content analyser (Operetta CLS Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

AbReview6938****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

ab18207 at 1/2000 dilution staining rat cerebellum tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). The tissue was formaldehyde fixed and a heat mediated antigen retrieval step was performed prior to incubation with the antibody for 16 hours. A biotinylated goat polyclonal antibody was used as the secondary.

This image is courtesy of Carl Hobbs, King's College London, United Kingdom

• WB

Lab

Western blot - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : Beta III Tubulin knockout HAP1 whole cell lysate (20 μg)

Lanes 1 - 2 : Merged signal (red and green). Green - ab18207 observed at 55 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab18207 was shown to recognize beta III Tubulin in wild-type HAP1 cells as signal was lost in beta III Tubulin knockout cells. An additional cross-reactive band at 50 kDa was observed in wild-type and knockout cells. Due to the immunogen's homology with TUB (Tubby protein homolog, Uniprot : P50607), this lower band could correspond to the TUB protein. Please note that cross-reactivity with this protein has not been confirmed experimentally.

Wild-type and beta III Tubulin knockout samples were subjected to SDS-PAGE. ab18207 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature

All lanes:

Western blot - Anti-beta III Tubulin antibody - Neuronal Marker (ab18207)

Predicted band size: 50 kDa

false

• WB

Project6840****

Western blot - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

All lanes:

Western blot - Anti-beta III Tubulin antibody - Neuronal Marker (ab18207) at 1 µg/mL

Lanes 1 and 4:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 10 µg

Lanes 2 and 5:

Brain (Mouse) Tissue Lysate at 10 µg

Lanes 3 and 6:

Brain (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 50 kDa

Observed band size: 55 kDa

false

Exposure time: 30s

• WB

Lab

Western blot - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab18207 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody , and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-beta III Tubulin antibody - Neuronal Marker (ab18207) at 1 µg/mL

Lane 1:

Brain (Mouse) Tissue Lysate at 10 µg

Lane 2:

Brain (Rat) Tissue Lysate at 10 µg

Lane 3:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 10 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 1/50000 dilution

Predicted band size: 50 kDa

Observed band size: 55 kDa

true

Exposure time: 30s

• WB

AbReview58465****

Western blot - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

All lanes:

Western blot - Anti-beta III Tubulin antibody - Neuronal Marker (ab18207) at 1/1000 dilution

All lanes:

Mouse hippocampus tissue lysate at 8 µg

Secondary

All lanes:

Goat anti-rabbit IgG (H&L) at 1/5000 dilution

Predicted band size: 50 kDa

Observed band size: 55 kDa

false

Exposure time: 10s

This image is courtesy of an abreview submitted by Dr Sergi Bayod.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

Immunohistochemistical staining (Formaldehyde/PFA-fixed paraffin-embedded sections) for Neuron specific beta III Tubulin antibody - Neuronal Marker (ab18207) on Dogfish/Catshark Tissue sections (head : snout region). Antigen retrieval step : Heat mediated. Blocking step : 1% BSA for 10 mins at RT. Primary Antibody used at 1/2000 incubated for 2 hours at RT. Secondary Antibody : Biotin labelled goat anti rabbit IgG (1/300).

This image is courtesy of Carl Hobbs, King's College London, United Kingdom