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

製品の概要

  • 製品名
    Anti-beta III Tubulin antibody [2G10] - Neuronal Marker
    beta III Tubulin 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [2G10] to beta III Tubulin - Neuronal Marker
  • 由来種
    Mouse
  • アプリケーション
    適用あり: Flow Cyt, IHC (PFA fixed), IHC-FoFr, ICC/IF, IHC-P, IHC-P, IP, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Rabbit, Chicken, Cow, Cat, Human, Quail, Common marmoset, Dogfish, Catshark
  • 免疫原

    Synthetic peptide corresponding to Rat beta III Tubulin aa 400-500.

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following lysates: SHSY-5Y whole cell, Human brain tissue, Human spinal cord tissue, SK N BE whole cell, mouse brain tissue, rat brain tissue IHC: Rat Cerebellum (FFPE tissue), Human cerebellum (FFPE) ICC/IF: NGF-differentiated PC12 cells
  • 特記事項

    This antibody clone [2G10] is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab78078 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
Flow Cyt Use 1µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IHC (PFA fixed) Use at an assay dependent concentration.
IHC-FoFr 1/300.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 0.5 µg/ml.
IHC-P Use a concentration of 0.2 - 1 µg/ml.
IP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).

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

画像

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Beta III Tubulin knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: Hek293 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab78078 observed at 50 kDa. Red - 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 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 ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • 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.

    See Abreview

  • 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.

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

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : 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: 90 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 ab78078 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution ab133406

  • 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) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1](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.

  • ab78078 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 ab78078 at 1μg/ml and ab6046 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Mouse secondary (ab150117) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Rabbit secondary (ab150088) 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 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.

  • IHC-FoFr image of Beta III tubulin (ab78078) staining on rat hippocamus sections. The sections used came from animals perfused fixed with Paraformaldehyde 4%, in phosphate buffer 0.2M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Brains were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.

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  • 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.
  • 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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  • 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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  • IHC image of ab78078 staining in mouse brain formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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.
  • 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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  • 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 70oC; 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.
  • 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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参考文献

This product has been referenced in:
  • Hao Q  et al. Procedure for the Isolation of Endothelial Cells from Human Cerebral Arteriovenous Malformation (cAVM) Tissues. Front Cell Neurosci 12:30 (2018). Read more (PubMed: 29467624) »
  • Ramanathan M  et al. RNA-protein interaction detection in living cells. Nat Methods 15:207-212 (2018). Read more (PubMed: 29400715) »

See all 53 Publications for this product

レビューと Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (brain)
Permeabilization
No
Specification
brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Formaldehyde
Username

Mr. Tamas Bellak

Verified customer

投稿 Mar 19 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Hippocampus)
Gel Running Conditions
Reduced Denaturing (13%)
Loading amount
10 µg
Specification
Hippocampus
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Dr. Sergi Bayod

Verified customer

投稿 Jan 05 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (DRG sensory neurons)
Permeabilization
Yes - 0.1% Triton X-100
Specification
DRG sensory neurons
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Sep 26 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (Ear base)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6
Permeabilization
No
Specification
Ear base
Blocking step
Protein Block serum free from Dako as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 37°C
Fixative
Formaldehyde
Username

Dr. Harshini Sarojini

Verified customer

投稿 Nov 17 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (differentiated neural stem cells)
Permeabilization
Yes - 0.2% Triton X 100
Specification
differentiated neural stem cells
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 May 06 2015

Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse neural stem cell in vitro differentiated for)
Gel Running Conditions
Reduced Denaturing (10% SDS PAGE)
Loading amount
1e+006 cells
Specification
mouse neural stem cell in vitro differentiated for
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C
Username

Abcam user community

Verified customer

投稿 Apr 30 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Sample
Mouse Tissue sections (Adult spinal cord, 20 micron)
Specification
Adult spinal cord, 20 micron
Permeabilization
Yes - Triton X-100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Mar 07 2015

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH 6
Sample
Mouse Tissue sections (Embryonic brain, 14 days, 16 micron)
Specification
Embryonic brain, 14 days, 16 micron
Permeabilization
Yes - Triton X-100
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

投稿 Dec 08 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Sample
Human Cell (human differentiated neuron)
Specification
human differentiated neuron
Permeabilization
Yes - 0.25% Triton-X
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

投稿 Nov 17 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (iPSC and neuronal differentiated cells)
Specification
iPSC and neuronal differentiated cells
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

投稿 Aug 06 2014

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