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RabMAb

Anti-beta III Tubulin抗体[EP1569Y] (ab52623)

概述

性能

应用

Our Abpromise guarantee covers the use of ab52623 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
IHC-Fr 1/100.
ICC/IF 1/500.
WB 1/1000 - 1/10000. Detects a band of approximately 52 kDa.
IP 1/40.
Flow Cyt 1/10 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/50 - 1/100.

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

图片

  • 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 - ab52623 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.

     

    Ab52623 was shown to specifically react with beta III Tubulin in wild-type cells as signal was lost in beta III Tubulin knockout HAP1 cells. Wild-type and beta III Tubulin knockout samples were subjected to SDS-PAGE. Ab52623 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

  • ab52623 staining beta III Tubulin in wild-type HAP1 cells (top panel) and TUBB3 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 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 with ab52623 at 1/500 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • ab52623 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 ab52623 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 [EP1569Y] (ab52623) at 1/1000 dilution

    Lane 1 : Brain (Mouse) Tissue Lysate
    Lane 2 : Brain (Rat) Tissue Lysate
    Lane 3 : Spinal Cord (Mouse) Tissue Lysate
    Lane 4 : Spinal Cord (Rat) Tissue Lysate
    Lane 5 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) at 1/10000 dilution

    Observed band size : 52 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 Licor blocking buffer before being incubated with ab52623 overnight at 4°C. Antibody binding was detected using ab175781 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

  • IHC-Fr image of Beta III Tubulin staining on zebrafish retina sections using ab52623 (1:100). The sections were paraformaldehyde and permeabilized using Triton-X. Antigen retrieval was performed using Tris-HCl and blocking was perfomed using 5% BSA for 1 hour at 23°C. ab38738 was diluted 1:100 and incubated with the sections for 16 hours at 4°C. The secondary antibody was goat polyclonal to rabbit IgG conjugated to Alexa Fluor 488 (1:1000).

    See Abreview

  • Overlay histogram showing U-87MG cells stained with ab52623 (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 (ab52623, 1/1000 dilution) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730, 0.1μg/1x106) 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-Tween for 20 min used under the same conditions.

  • Anti-beta III Tubulin antibody [EP1569Y] (ab52623) at 1/10000 dilution + HeLa cell lysate at 10 µg

    Secondary
    goat anti-rabbit HRP at 1/2000 dilution

    Observed band size : 52 kDa (why is the actual band size different from the predicted?)
  • ab52623, at a 1/50 dilution, staining human class III beta Tubulin in breast carcinoma tissue using Immunohistochemistry, Paraffin embedded tissue.
  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling beta III Tubulin with ab52623 at 1/20 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

文献

This product has been referenced in:
  • Zhang H  et al. Paeoniflorin Ameliorates Experimental Autoimmune Encephalomyelitis via Inhibition of Dendritic Cell Function and Th17 Cell Differentiation. Sci Rep 7:41887 (2017). WB ; Mouse . Read more (PubMed: 28165507) »
  • Eng GWL  et al. Site-specific phosphorylation of casein kinase 1 d (CK1d) regulates its activity towards the circadian regulator PER2. PLoS One 12:e0177834 (2017). Read more (PubMed: 28545154) »

See all 10 Publications for this product

客户评价及客户问答

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mm sodium citrate PH6
Permeabilization
No
Specification
tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Oct 27 2017

Application
Western blot
Sample
Rat Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions
Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount
14 µg
Specification
Intestinal tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Julien Chevalier

Verified customer

提交于 Sep 30 2015

Application
Western blot
Sample
Human Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions
Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount
14 µg
Specification
Intestinal tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Julien Chevalier

Verified customer

提交于 Sep 30 2015

Application
Western blot
Sample
Mouse Tissue lysate - whole (Intestinal tissue)
Gel Running Conditions
Reduced Denaturing (4-12% Bis Tris MOPS)
Loading amount
14 µg
Specification
Intestinal tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Julien Chevalier

Verified customer

提交于 Sep 30 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Cryopreserved embryonic cortical neurons (QBMcells)
Specification
Cryopreserved embryonic cortical neurons (QBMcells
Permeabilization
No
Fixative
paraformaldehyde with picric acid
Username

Ms. Babben Tinner

Verified customer

提交于 Feb 26 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Sample
Zebrafish Tissue sections (Retina, Inner plexiform layer)
Specification
Retina, Inner plexiform layer
Permeabilization
Yes - triton X
Fixative
Paraformaldehyde
Username

Dr. Ryan Macdonald

Verified customer

提交于 Aug 22 2014

Thank you once again for your enquiry and for your patience. I am sorry to confirm that for Beta tubulin, the antibodies we have available tested and guaranteed in EM and mouse, rat and human tissues are mouse monoclonals. I can suggest to consi...

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