製品の概要

  • 製品名Anti-beta III Tubulin antibody
    beta III Tubulin 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to beta III Tubulin
  • 特異性The immunising sequence is found in both beta III and beta IV tubulin. ab18207 detects human neuron specific beta III Tubulin protein specifically and cleanly but not as strongly as it detects the equivalent mouse and rat protein.
  • アプリケーション適用あり: IHC-FoFr, Flow Cyt, IHC - Wholemount, IHC-P, IHC-P, IHC-Fr, ICC/IF, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Human, Pig, Marmoset (common), Dogfish/Catshark
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human neuron specific beta III Tubulin.

    (Peptide available as ab18660.)

  • ポジティブ・コントロール
    • Human, Mouse, Rat or Dogfish/Catshark Brain Tissue ICC/IF: SKNSH, Neuro-2A and NGF-differentiated PC12 cells IHC: Hu Cerebellum (FFPE tissue) Flow Cytometry: U-87MG cells, Neuro 2A cells.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab18207 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
IHC-FoFr Use at an assay dependent concentration. PubMed: 20568963
Flow Cyt Use 0.01µg for 106 cells.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC - Wholemount Use at an assay dependent concentration. PubMed: 25383879
IHC-P 1/2000.
IHC-P 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 - 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 50-55 kDa (predicted molecular weight: 50 kDa).Can be blocked with Human beta III Tubulin peptide (ab18660).

ターゲット情報

  • 機能Tubulin is the major constituent of microtubules. It binds two moles of GTP, one at an exchangeable site on the beta chain and one at a non-exchangeable site on the alpha-chain. TUBB3 plays a critical role in proper axon guidance and mantainance.
  • 組織特異性Expression is primarily restricted to central and peripheral nervous system.
  • 関連疾患Defects in TUBB3 are the cause of congenital fibrosis of extraocular muscles type 3A (CFEOM3A) [MIM:600638]. A congenital ocular motility disorder marked by restrictive ophthalmoplegia affecting extraocular muscles innervated by the oculomotor and/or trochlear nerves. It is clinically characterized by anchoring of the eyes in downward gaze, ptosis, and backward tilt of the head. Congenital fibrosis of extraocular muscles type 3 presents as a non-progressive, autosomal dominant disorder with variable expression. Patients may be bilaterally or unilaterally affected, and their oculo-motility defects range from complete ophthalmoplegia (with the eyes fixed in a hypo- and exotropic position), to mild asymptomatic restrictions of ocular movement. Ptosis, refractive error, amblyopia, and compensatory head positions are associated with the more severe forms of the disorder. In some cases the ocular phenotype is accompanied by additional features including developmental delay, corpus callosum agenesis, basal ganglia dysmorphism, facial weakness, polyneuropathy.
  • 配列類似性Belongs to the tubulin family.
  • ドメインThe highly acidic C-terminal region may bind cations such as calcium.
  • 翻訳後修飾Some glutamate residues at the C-terminus are polyglutamylated. This modification occurs exclusively on glutamate residues and results in polyglutamate chains on the gamma-carboxyl group. Also monoglycylated but not polyglycylated due to the absence of functional TTLL10 in human. Monoglycylation is mainly limited to tubulin incorporated into axonemes (cilia and flagella) whereas glutamylation is prevalent in neuronal cells, centrioles, axonemes, and the mitotic spindle. Both modifications can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of such modifications is still unclear but they regulate the assembly and dynamics of axonemal microtubules.
  • 細胞内局在Cytoplasm > cytoskeleton.
  • Information by UniProt
  • 参照データベース
  • 別名
    • beta 3 tubulin antibody
    • beta-4 antibody
    • CDCBM antibody
    • CDCBM1 antibody
    • CFEOM3 antibody
    • CFEOM3A antibody
    • FEOM3 antibody
    • M(beta)3 antibody
    • M(beta)6 antibody
    • MC1R antibody
    • Neuron specific beta III Tubulin antibody
    • Neuron-specific class III beta-tubulin antibody
    • QccE-11995 antibody
    • QccE-15186 antibody
    • TBB3_HUMAN antibody
    • Tubb 3 antibody
    • TUBB3 antibody
    • TUBB4 antibody
    • Tubulin beta 3 antibody
    • Tubulin beta 3 chain antibody
    • Tubulin beta 4 antibody
    • Tubulin beta III antibody
    • Tubulin beta-3 chain antibody
    • Tubulin beta-4 chain antibody
    • Tubulin beta-III antibody
    see all

Anti-beta III Tubulin antibody 画像

  • 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.

  • ab18207 staining beta III Tubulin in NGF-differentiated PC12 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 5μg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Rabbit secondary (ab150081) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Mouse secondary (ab150120) 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.

  • All lanes : Anti-beta III Tubulin antibody (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
    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 kDa (why 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 an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

  • IHC image of ab18207 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 (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab18207, 1μg/ml 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.

  • 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 an AlexaFluor®488 Goat anti-Rabbit secondary (ab150081) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Mouse secondary (ab150120) 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.

  • All lanes : Anti-beta III Tubulin antibody (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
    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 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds
  • 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).
  • ab18207 staining beta III Tubulin in SKNSH 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 an AlexaFluor®488 Goat anti-Rabbit secondary (ab150081) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Mouse secondary (ab150120) 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.

  • 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 Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (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.

  • Overlay histogram showing U-87MG 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 Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/4000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab171870, 0.01μg/1x10cells) 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.

  • 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.

    See Abreview

  • ab18207 staining beta III tubulin in PC-12 cells treated with venlafaxine hydrochloride (ab120715), by ICC/IF. Increase in the number and length of neurites (stained with beta III tubulin) correlates with increased concentration of venlafaxine hydrochloride, as described in literature.
    The NGF treated cells were incubated at 37°C for 6 hour in media containing different concentrations of ab120715 (venlafaxine hydrochloride) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab18207 (1 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody.
  • ICC/IF image of ab18207 stained SH-SY-5Y cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab18207, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • ab18207 staining human cultured stem cells differentiated to neurons, by ICC/IF.  Cells were PFA fixed and permeabilized in Triton X prior to blocking with 1% BSA for 30 minutes at 27°C.  The primary antibody was used undiluted and incubated with the sample for 24 hours at 4°C.  A PE-conjugated rabbit anti-rabbit antibody was used as the secondary.

    See Abreview

  • 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.

    See Abreview

  • ab18207 staining rat somatosensory cortex tissue sections by IHC-Fr.  Sections were PFA fixed and permeabilized in TritonX100 prior to blocking in 3% BSA for 1 hour at RT.  The primary antibody was diluted 1/1000 and incubated with the sample for 18 hrs at 4°C.  A biotinylated goat anti-rabbit IgG was used as the secondary antibody. The anti-beta III tubulin staining is clearly visible in dendrites of pyramidal cells passing from layer V to the superficial layers.  (Magnification = 200x)

    See Abreview

Anti-beta III Tubulin antibody (ab18207) 使用論文

This product has been referenced in:
  • Long K  et al. Integrin signalling regulates the expansion of neuroepithelial progenitors and neurogenesis via Wnt7a and Decorin. Nat Commun 7:10354 (2016). IHC-Fr, IHC-FrFl ; Mouse . Read more (PubMed: 26838601) »
  • Ren M  et al. A biofidelic 3D culture model to study the development of brain cellular systems. Sci Rep 6:24953 (2016). ICC/IF ; Rat . Read more (PubMed: 27112667) »

See all 44 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Neuron)
Permeabilization Yes - tritonx100
Specification Neuron
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative Paraformaldehyde
Username

Ms. kyungjoo seong

Verified customer

投稿 Nov 02 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Penguin Tissue sections (brain)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: tris-EDTA, pH9.0
Permeabilization No
Specification brain
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

投稿 Nov 01 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Fukomys anselli Tissue sections (Retina)
Antigen retrieval step None
Permeabilization Yes - 0.2% Triton
Specification Retina
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 6% · Temperature: RT°C
Fixative Paraformaldehyde
Username

Dr. Pascal Malkemper

Verified customer

投稿 Jul 04 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Glioma)
Gel Running Conditions Reduced Denaturing (12% gel)
Loading amount 25 µg
Specification Glioma
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Apr 27 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Finch (Fringillidae) Tissue sections (brain)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization No
Specification brain
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

投稿 Mar 23 2016

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (brain)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization No
Specification brain
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Jan 29 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (U87 (Human glioblastoma cells))
Permeabilization Yes - 0.2% TritonX-100 in 1xPBS for 15min in room temperature.
Specification U87 (Human glioblastoma cells)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 24°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Dec 24 2015

Application Western blot
Sample Mouse Cell lysate - whole cell (Neuro-2a)
Gel Running Conditions Reduced Denaturing (10%)
Loading amount 40 µg
Treatment Serum deprivation for 24h
Specification Neuro-2a
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C
Username

Mr. gajendra reddy

Verified customer

投稿 Nov 02 2015

Application Immunocytochemistry
Sample Human Cell (NEURONAL CELL)
Permeabilization Yes - TRITON X-100
Specification NEURONAL CELL
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.5% · Temperature: RT°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

投稿 Sep 30 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (nerve)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization No
Specification nerve
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

投稿 Sep 28 2015

1-10 of 48 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"