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

製品の概要

  • 製品名Anti-beta III Tubulin antibody [2G10]
    beta III Tubulin 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [2G10] to beta III Tubulin
  • 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, Marmoset (common), Dogfish/Catshark
  • 免疫原

    Tissue, cells or virus corresponding to Rat beta III Tubulin.

  • 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

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

Anti-beta III Tubulin antibody [2G10] 画像

  • 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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  • 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] (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
    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.

  • Immunohistochemistical detection of Neuron specific beta III Tubulin using antibody ab78078 on PFA fixed mouse embryo (image shows cervical spinal cord T/S). Primary Antibody Dilution: 1/1000 for 1 hour in TBS/BSA/azide. Secondary Antibody: anti Mouse IgG conjugated to Alexa Fluor® 594 (1/1000). The image submiteed here shows cervical spinal cord before closure.

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  • 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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  • ab78078 staining a p8 mouse RMS neuroblast explant embedded in a 3D Matrigel matrix by ICC/IF. The explant was fixed with 4% paraformaldehyde for 45 minutes at room temperature and permeabilized with 0.3%Triton X.  The explant was then blocked with 15% goat serum for 30 minutes at 21°C. The cultured explant was then stained with ab78078 at 1/2000 in 0.3% TritonX with 0.1x PBS and 15% goat serum for 16h at 4°C. An Alexa Fluro 488 goat anti-mouse polyclonal antibody at 1/1000 was used as the secondary antibody. Beta III tubulin (which stains the axons) can be observed in green and the nuclei can be obbserved in blue (DAPI).

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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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  • ICC/IF image of ab78078 stained PC12 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab78078, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) PC12 cells at 5µg/ml.
  • 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.
  • 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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  • ab78078 staining Neuron specific beta III Tubulin in human colon tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval in Citric acid pH6 and then blocking with 1% BSA was performed for 10 minutes at RT. The primary antibody was used at dilution at 1/200 and incubated with sample in TBS/BSA/azide for 2 hours. A Biotin conjugated goat polyclonal to mouse IgG was used at dilution at 1/200 as secondary antibody. Images A and B represent staining of the clone TU20 and 2G10 respectively in ganglia of Auerbach's plexus.

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Anti-beta III Tubulin antibody [2G10] (ab78078) 使用論文

This product has been referenced in:
  • Heeren AM  et al. On the development of extragonadal and gonadal human germ cells. Biol Open 5:185-94 (2016). Read more (PubMed: 26834021) »
  • Iordache F  et al. Electrophysiology, immunophenotype, and gene expression characterization of senescent and cryopreserved human amniotic fluid stem cells. J Physiol Sci 66:463-476 (2016). Read more (PubMed: 27053101) »

See all 23 Publications for this product

Product Wall

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
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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
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Dr. Harshini Sarojini

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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
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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
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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
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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
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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
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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
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投稿 Aug 06 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Sample Rat Cell (neuroblast)
Specification neuroblast
Permeabilization Yes - 0.2% triton
Fixative Formaldehyde
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投稿 Sep 26 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citric buffer, pH6.0
Sample Human Tissue sections (adult skin)
Specification adult skin
Permeabilization Yes - Tween-20
Fixative Paraformaldehyde
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Mr. Heiko Locher

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投稿 Sep 05 2013

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