Anti-beta III Tubulin抗体[TU-20] (ab7751)

概述

  • 产品名称Anti-beta III Tubulin抗体[TU-20]
    参阅全部 beta III Tubulin 一抗
  • 描述
    小鼠单克隆抗体[TU-20] to beta III Tubulin
  • 特异性Class III beta-tubulin specific for neurones.
  • 经测试应用适用于: ICC, IHC-P, WB, IHC-FoFr, IHC-Fr, ICC/IF, IHC-P, IP, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Chicken, Hamster, Cow, Cat, Human, Pig, Carp, Baboon
    预测可用于: a wide range of other species, all Mammals
  • 免疫原

    Synthetic peptide corresponding to Human beta III Tubulin aa 441-448 conjugated to Keyhole Limpet Haemocyanin (KLH).
    Sequence:

    ESESQGPK

  • 表位ESESQGPK (aa 441-448)
  • 阳性对照
    • ICC/IF: Neuro-2a cells IHC: Hu Cerebellum (FFPE tissue)

性能

应用

Our Abpromise guarantee covers the use of ab7751 in the following tested applications.

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

应用 Ab评论 说明
ICC 1/500.
IHC-P Use a concentration of 10 µg/ml. Pretreatment: 0.1% pepsin (trypsin) in 0.1 M HCl for 30 minutes at RT. High-temperature citrate buffer antigen retrieval can also be performed.
WB Use a concentration of 1 - 2 µg/ml. We suggest reducing conditions and a 90 minute incubation time.
IHC-FoFr Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC/IF 1/1000.
IHC-P Use a concentration of 5 µg/ml.
IP Use at an assay dependent concentration.
Flow Cyt 1/20 - 1/50.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

靶标

  • 功能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.
  • 组织特异性Expression is primarily restricted to central and peripheral nervous system.
  • 疾病相关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.
  • 序列相似性Belongs to the tubulin family.
  • 结构域The highly acidic C-terminal region may bind cations such as calcium.
  • 翻译后修饰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.
  • 细胞定位Cytoplasm > cytoskeleton.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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 [TU-20] 图像

  • All lanes : Anti-beta III Tubulin antibody [TU-20] (ab7751) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466) at 20 µg
    Lane 2 : Human spinal cord tissue lysate - total protein (ab29188)

    Secondary
    IRDye® 800CW Goat Anti-Mouse at 1/10000 dilution

    Performed under reducing conditions.

    Observed band size : 51 kDa (why is the actual band size different from the predicted?)

    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 LI-COR® blocking buffer before being incubated with ab7751 overnight at 4°C. Antibody binding was detected using the IRDye® 800CW Goat Anti-Mouse secondary at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Odyssey® CLx Imaging System.

  • IHC image of ab7751 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 ab7751, 5μg/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.

  • ab7751 staining beta III Tubulin in induced Neuron derived from Human pluripotent cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.25% Triton X-100 in PBS and blocked with 1% BSA for 30 minutes at 22°C. Samples were incubated with primary antibody (1/500) for 16 hours at 4°C. An Alexa Fluor® 594-conjugated Donkey anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • Overlay histogram showing SH-SY5Y cells stained with ab7751 (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 (ab7751, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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 (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • ab7751 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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  • ab7751 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 ab7751 at 1/1000 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.

  • Immunohistochemistical detection of Neuron specific beta III Tubulin using antibody (ab7751) on whole mouse e12 embryo (formaldehyde fixed/frozen sections). Permeabilization: No. Blocking step: 1% BSA for 10 mins @ rt°C. Primary Antibody Dilution 1/200; Incubation time 2 hours in TBS/BSA/azide. Secondary antibody: anti Mouse IgG Conjugated to Alexa Fluor® 594 (1/1000). Floorplate region of developing cervical spinal cord (I have not studied this particular region).

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  • ab7751 immunostaining (1/500) neuronal processes in primary mouse cortical culture.

  • Immunohistochemical detection with Neuron specific beta III Tubulin antibody [TU-20] (ab7751) on formaldehyde fixed rat spinal cord tissue sections. Antigen retrieval step: heat mediated in citric acid pH6. Blocking: 1% BSA for 10 mins@ rt°C. Primary antibody ab7751 incubated at 1/500 for 2 hours. Secondary antibody: anti mouse IgGs conjugated to biotin (1/200).  ab7751 shows good fibrillar positivity in rat spinal motor neurone cell bodies: see image. Some L/S nerve fibre processes are clearly seen yet the majority in the white matter are also not obvious.

    See Abreview

实验方案

Anti-beta III Tubulin antibody [TU-20] (ab7751)参考文献

This product has been referenced in:
  • Bryukhovetskiy I  et al. Cancer stem cells and microglia in the processes of glioblastoma multiforme invasive growth. Oncol Lett 12:1721-1728 (2016). Rat . Read more (PubMed: 27602106) »
  • Laprairie RB  et al. The cytokine and endocannabinoid systems are co-regulated by NF-?B p65/RelA in cell culture and transgenic mouse models of Huntington's disease and in striatal tissue from Huntington's disease patients. J Neuroimmunol 267:61-72 (2014). Mouse, Human . Read more (PubMed: 24360910) »

See all 31 Publications for this product

Product Wall

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 (8%)
Loading amount 5 µg
Specification Hippocampus
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Sergi Bayod

Verified customer

提交于 Aug 03 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (induced Neuron derived from pluripotent cells)
Permeabilization Yes - 0.25% Triton X-100 in PBS
Specification induced Neuron derived from pluripotent cells
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Dec 16 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (differentiated cultured mouse neural stem cells)
Permeabilization Yes - 0.2% triton x 100 in PBS
Specification differentiated cultured mouse neural stem cells
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Apr 29 2015

Application Western blot
Sample Mouse Cell lysate - whole cell (in vitro differentiated neural stem cell)
Gel Running Conditions Reduced Denaturing (10% SDS PAGE)
Loading amount 5 µg
Specification in vitro differentiated neural stem cell
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Apr 29 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Sample Mouse Cell (primary cortical neuron)
Specification primary cortical neuron
Permeabilization Yes - 0.1% Triton/1% BSA/PBS
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Jun 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 30 minute(s) · Concentration: 5% · Temperature: RT°C
Sample Mouse Cell (Dorsal root ganglion culture)
Specification Dorsal root ganglion culture
Permeabilization Yes - Methanol
Fixative Paraformaldehyde
Username

Mr. Heiko Locher

Verified customer

提交于 Sep 23 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 500000 cells
Gel Running Conditions Non-reduced Denaturing (4-12% Bis Tris gel)
Sample Human Cell lysate - whole cell (SH-SY5Y)
Specification SH-SY5Y
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Aug 20 2013

Thank you for contacting us.

I think either of the two beta III tubulin clones will be effective, but I would choose ab7751 (the TU-20 clone) simply because it has been published in more references than the other clone, ab78078 (2G10 clone)....

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Glioblastoma)
Specification Glioblastoma
Fixative Paraformaldehyde
Permeabilization Yes - 1% Triton X-100
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Jul 04 2012

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab98787.

To check the status of the order please contact o...

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