Anti-beta III Tubulin抗体- Neuronal Marker
参阅全部 beta III Tubulin 一抗
兔多克隆抗体to beta III Tubulin - Neuronal Marker
Rabbit
The immunogen used for this product shares 75% homology with TUB (Tubby protein homolog, Uniprot: P50607). In western blot, we observe a specific band at ~55kDa which is not seen in KO cell lines. An additional band below this band of interest is seen at ~50kDa in both the WT and KO cells which could correspond to the protein TUB. Please note that cross-reactivity with this protein has not been confirmed experimentally. TUB is localized notably in high concentrations in the nucleoli of brain neurons with lower protein levels in the cytoplasm. Please, therefore, be aware that ICC experiments may need to be optimised. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above. As an alternative antibody, we would recommend our recombinant rabbit monoclonal antibody ab52623 which has been shown to specific in both WB and ICC using KO cells.
适用于: ICC/IF, Flow Cyt (Intra), IHC-P, WBmore details
与反应: Mouse, Rat, Human, Common marmoset, Dogfish, Catshark
预测可用于: Pig, Rhesus monkey
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available as ab18660)
Flow Cyt (Intra): Neuro 2A, U-87MG cells. IHC-P: Rat cerebellum, mouse brain, adult mouse ovaries, Dogfish/Catshark tissue (snout region). ICC/IF: SK-N-SH cells, Neuro 2A, PC12 and RA induced P19 cells. Primary rat neurons/glia, DIV14. WB: WT HAP-1 whole cell lysate. Human, mouse and rat brain tissue lysate. Mouse hippocampus tissue lysate.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab18207于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| ICC/IF | (19) | Use a concentration of 1 µg/ml. |
| Flow Cyt (Intra) | Use 0.01µg for 106 cells. ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. We recommend Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody. | |
| IHC-P | (17) | 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
| WB | (10) | Use a concentration of 1 µg/ml. Detects a band of approximately 50-55 kDa (predicted molecular weight: 50 kDa). |
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Unigene: 40068 Mouse
Unigene: 43958 Rat
Entrez Gene: 10381 Human
Entrez Gene: 22152 Mouse
Omim: 602661 Human
SwissProt: Q13509 Human
SwissProt: Q9ERD7 Mouse
Unigene: 511743 Human
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M(beta)6 antibody
MC1R antibody
Neuron specific beta III Tubulin antibody
Neuron-specific class III beta-tubulin antibody
QccE-11995 antibody
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TBB3_HUMAN antibody
Tubb 3 antibody
TUBB3 antibody
TUBB4 antibody
Tubulin beta 3 antibody
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tuj 1 antibody
tuj1 antibody
beta 3 tubulin antibody
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beta-4 antibody
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 0.32µg/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.
Immunocytochemistry - Anti-beta III Tubulin antibody - Neuronal Marker (ab18207)This image is courtesy of an Abreview submitted by Lenin Veeraval
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.
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.
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.
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
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.
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 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 22ºC. The secondary antibody used was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) at 1/4000 dilution for 30 min at 22ºC. 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 >5,000 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.
Western blot - Anti-beta III Tubulin antibody - Neuronal Marker (ab18207)
All lanes : Anti-beta III Tubulin antibody - Neuronal Marker (ab18207) at 1 µg/ml
Lane 1 : Human brain tissue lysate - total protein (ab29466)
Lane 2 : Brain (Mouse) Tissue Lysate
Lane 3 : Brain (Rat) Tissue Lysate
Lane 4 : Human brain tissue lysate - total protein (ab29466) with Human beta III Tubulin peptide (ab18660) at 2 µg/ml
Lane 5 : Brain (Mouse) Tissue Lysate with Human beta III Tubulin peptide (ab18660) at 2 µg/ml
Lane 6 : Brain (Rat) Tissue Lysate with Human beta III Tubulin peptide (ab18660) at 2 µg/ml
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 55 kDawhy is the actual band size different from the predicted?
Exposure time: 30 seconds
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody - Neuronal Marker (ab18207)This image is courtesy of Carl Hobbs, King's College London, United Kingdom
ab18207 at 1/2000 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.
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 22ºC. The secondary antibody used was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) at 1/4000 dilution for 30 min at 22ºC. 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 >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Western blot - Anti-beta III Tubulin antibody - Neuronal Marker (ab18207)
All lanes : Anti-beta III Tubulin antibody - Neuronal Marker (ab18207) at 1 µg/ml
Lane 1 : Brain (Mouse) Tissue Lysate
Lane 2 : Brain (Rat) Tissue Lysate
Lane 3 : Human brain tissue lysate - total protein (ab29466)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 55 kDawhy is the actual band size different from the predicted?
Exposure time: 30 seconds
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody - Neuronal Marker (ab18207)This image is courtesy of Carl Hobbs, King's College London, United Kingdom
ab18207 at 1/2000 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody - Neuronal Marker (ab18207)This image is courtesy of Carl Hobbs, King's College London, United Kingdom
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).
Western blot - Anti-beta III Tubulin antibody - Neuronal Marker (ab18207)This image is courtesy of an abreview submitted by Dr Sergi Bayod.
All lanes : Anti-beta III Tubulin antibody - Neuronal Marker (ab18207) at 1/1000 dilution
All lanes : Mouse hippocampus tissue lysate
Lysates/proteins at 8 µg per lane.
Secondary
All lanes : Goat anti-rabbit IgG (H&L) at 1/5000 dilution
Predicted band size: 50 kDa
Observed band size: 55 kDawhy is the actual band size different from the predicted?
Exposure time: 10 seconds
Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody - Neuronal Marker (ab18207)Sheikh, M.A. et al PLoS One. 2013;8(2):e55826. doi: 10.1371/journal.pone.0055826. Epub 2013 Feb 7 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Differential expression of Dnmt1, Dnmt3a, and Dnmt3b during RA induced neuronal differentiation of P19 cells
Mouse P19 cells either left untreated (top panel) or RA treated for initial 2 days and further cultured for 4 days without RA (6 days, bottom panel) were immunostained with neuron specific β-III tubulin antibody and nuclei were stained using DAPI.
In order to confirm the neuronal morphology, the cells were stained for neuron specific beta III-tubulin (ab18207). RA induced P19 cells showed immunoreactivity against βIII-tubulin, indicating a neuronal phenotype. In contrast, undifferentiated P19 cells were βIII-tubulin negative.
(After Figure 1A of Sheikh et al)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody - Neuronal Marker (ab18207)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/.
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.
