Anti-beta III Tubulin antibody [2G10] - Neuronal Marker

16 product images

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  Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  ab78078 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 ab78078 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) 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.

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  Grimaldi, A. et al PLoS One. 2015 Dec 15;10(12):e0145018. doi: 10.1371/journal.pone.0145018. eCollection 2015. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  Soft palate innervation is primarily oro-medial and complements the differentiated muscle pattern

  (A-L) MHC (green) and β3-tubulin (red) co-immunostaining along the antero-posterior axis of the mouse soft palate at E13.5 (A-C), E14.5 (D-F), E16.5 (G-I), and P0 (J-L) at the level of the TVP and PLG (A, D, G, J), LVP (B, E, H, K), and PLP and SPC (C, F, I, L). Dashed lines indicate the outline of the palatal shelf and tongue epithelium. Arrows indicate the nerve fibers innervating the palatal shelves in a pattern complementary to the differentiated muscle, mainly located orally in the medial region. The schematic drawings indicate the orientation and the position of each section. Scale bars: 200μm.

  beta III Tubulin is detected with ab78078.

  (From Figure 4 of Grimaldi et al)

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

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  Immunohistochemistical detection of Neuron specific beta III Tubulin antibody [2G10] using ab78078 in formaldehyde-fixed paraffin-embedded quail embryo eye sections. Antigen retrieval step: heat mediated in citric acid pH6 buffer. Blocking step: 1% BSA for 10 mins @ rt°C Primary antibody incubated at 1/1250 for 2 hours in TBS/BSA/azide. Secondary antibody: anti-Mouse IgG conjugated to biotin used at 1/200. In this image, the lining cells of the hyaloid artery are positive. Excellent neural component specificity: even small fibres outside of the cartilage model of the orbit are clearly demonstrated.

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

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  Immunohistochemistical detection of Neuron specific beta III Tubulin using antibody (ab78078) on formaldehyde-fixed paraffin-embedded human medulla oblongata sections. Antigen retrieval step: Heat mediated in citric acid HIER buffer. Permeabilization: No. Blocking step: 1% BSA for 10 mins @ rt°C. Primary antibody dilution 1/250 incubated for 2 hours in TBS/BSA/azide. Secondary antibody: anti Mouse IgGs conjugated to biotin (1/200). This section was cut from an anonymous autopsy P.wax block that is over 20 yrs old, so I would expect variable positivity when compared with more recently sampled tissues (giving higher dilution factors, such as I obtained with fresh mouse/rat CNS blocks.) Transverse-cut axons stain very intensely longitudinal nerve processes and cell bodies are not so heavily stained but are still easily seen.

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  Immunoprecipitation - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  Neuron specific beta III Tubulin - Neuronal Marker was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 5µg of Mouse monoclonal [2G10] to Neuron specific beta III Tubulin - Neuronal Marker and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
  The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
  Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab78078.
  Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
  Band: 50kDa: Neuron specific beta III Tubulin - Neuronal Marker.

  All lanes: Immunoprecipitation - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  Predicted band size: 50 kDa

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  Western blot - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  Lanes 1 - 4: Merged signal (red and green). Green - ab78078 observed at 50 kDa. Red - loading control, Anti-GAPDH antibody [EPR16891] - Loading Control ab181602, observed at 37 kDa.
  

  ab78078 was shown to specifically react with beta III Tubulin in wild-type HAP1 cells as signal was lost in beta III Tubulin knockout cells. Wild-type and beta III Tubulin knockout samples were subjected to SDS-PAGE. ab78078 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 10 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078) at 1/20000 dilution

  Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

  Lane 2: Beta III Tubulin knockout HAP1 whole cell lysate at 20 µg

  Lane 3: HeLa whole cell lysate at 20 µg

  Lane 4: HEK-293 whole cell lysate at 20 µg

  Predicted band size: 50 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  ab78078 staining beta III Tubulin in NGF-differentiated PC12 cells. The cells were fixed with 100% methanol (5 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 ab78078 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) 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.

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  Western blot - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  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 ab78078 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406

  All lanes: Western blot - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078) at 1 µg/mL

  Lane 1: Human brain tissue lysate - total protein (ab29466) at 10 µg

  Lane 2: Brain (Mouse) Tissue Lysate at 10 µg

  Lane 3: Brain (Rat) Tissue Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (Goat Anti-Mouse IgG H&L (HRP) preadsorbed ab97040) at 1/50000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 50 kDa

  Observed band size: 50 kDa

  Exposure time: 90s

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  This image is courtesy of an anonymous customer review

  Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  ab78078 staining beta III Tubulin (red) in mouse differentiated neural stem cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.2% Triton X-100. Samples were incubated with primary antibody (5μg/ml in PBS + 3% BSA) for 16 hours at 4°C. An Alexa Fluor^® 568-conjugated donkey anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody. Blue - DAPI nuclear counterstain.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  IHC image of ab78078 staining beta III Tubulin in Human 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 (pH 6.0 epitope retrieval solution 1) for 20 mins. The section was then incubated with ab78078, 0.5μg/ml, 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.

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

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  Immunohistochemical detection of Neuron specific beta III Tubulin using ab78078 on formaldehyde fixed dogfish/catshark PNS tissue sections. Antigen retrieval step: heat mediated in citric acid. Blocking: 1% BSA for 10 mins @ rt°C. Primary antibody ab78078 incubated at 1/1250 for 2 hours. Secondary Antibody: anti mouse IgG conjugated to biotin @ 1/200 This is a composite image of neuronal cell bodies and fibres; the upper image uses coloured arrowheads to indicate positive PNS components: cell bodies (black), L/S (green) and T/S (red) nerve fibres of what appear to be three Ganglia. Note that the L/S fibres do not appear as well-stained as the T/S fibres. The lower image shows what appears to be a linear sequence of single nerve cells and their processes within the core of a gill.

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

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  Immunohistochemistical detection of Neuron specific beta III Tubulin antibody using ab78078 on formaldehyde-fixed paraffin-embedded marmoset cerebellum sections. Antigen retrieval step: Heat mediated in Citric acid pH6 buffer. Permeabilization: No. Blocking step: 1% BSA for 10 mins @ rt°C. Primary Antibody used at 1/1000 incubated for 2 hours in TBS/BSA/azide. Secondary Antibody: anti mouse IgG conjugation to biotin (1/200).

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  This image is courtesy of a customer review by Carl Hobbs

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  ab78078 staining beta III Tubulin in Chicken e6: developing eye tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at room temperature; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/1250) for 2 hours. A Biotin-conjugated Goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

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  Flow Cytometry (Intracellular) - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  Overlay histogram showing SH-SY5Y stained with ab78078 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween 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 (ab78078, 1µg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1](Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol/permeabilized in 0.1% PBS-Tween used under the same conditions.
  
  

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  IHC image of ab78078 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 EDTA/Tris (epitope retrieval solution 2) for 20 mins. The section was then incubated with ab78078, 0.5μg/ml, 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.
  
  

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [2G10] - Neuronal Marker (ab78078)

  IHC image of ab78078 staining in mouse brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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 20 mins. The section was then incubated with ab78078, 1µg/ml, 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.
  
  
  
  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.