Overview

  • Product name
    Anti-beta III Tubulin antibody [EP1569Y]
    See all beta III Tubulin primary antibodies
  • Description
    Rabbit monoclonal [EP1569Y] to beta III Tubulin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, WB, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Zebrafish
  • Immunogen

    Synthetic peptide within Human beta III Tubulin (C terminal). The exact sequence is proprietary.

  • Positive control
    • ICC/IF: PC-12 and wild-type HAP1 cells. WB: HAP1 cell lysate. HeLa, PC-12 and HEK-293 whole cell lysate. Mouse and rat brain and spinal cord tissue lysate. IHC-P: Human breast carcinoma tissue. Flow Cyt: U-87 MG and HeLa cells. IHC-Fr: Zebrafish retina sections.
  • General notes

    A trial size is available to purchase for this antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • Function
    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.
  • Tissue specificity
    Expression is primarily restricted to central and peripheral nervous system.
  • Involvement in disease
    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.
  • Sequence similarities
    Belongs to the tubulin family.
  • Domain
    The highly acidic C-terminal region may bind cations such as calcium.
  • Post-translational
    modifications
    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.
  • Cellular localization
    Cytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • 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 for 5 minutes, 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 1 hour. 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 1 hour with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green).

    Nuclear DNA was labeled in blue with DAPI.

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

  • 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.

  • In vivo, CK1δ T347 is phosphorylated in trans.

    D) Phosphorylation of CK1δ at T347 is not dependent on CK1 activity. HEK-293 cells over-expressing wild-type CK1δ were treated with CK1 inhibitors PF670462 (PF670, 1 μM), PF4800567 (PF480, 1 μM), or D4476 (30 μM) for 3 hours, followed by addition for 30 min of either DMSO or 40 nM Calyculin A. Cell lysates were immunoblotted for anti-pT347 or anti-Myc antibodies. CK1 inhibitors block the autophosphorylation-induced mobility shift but do not decrease phosphorylation of T347. Myc-tagged CK1δ indicated by curly brackets. ab52623 lower panel.

  • ab52623 staining beta III Tubulin in NGF-differentiated PC-12 (Rat adrenal gland pheochromocytoma cell line) cells.

    The cells were fixed with 100% methanol (5 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab52623 at 5 μg/ml and ab7291 at 1 µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1 hour 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 labeled 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.

  • ab52623, at a 1/50 dilution, staining class III beta Tubulin in human breast carcinoma tissue using Immunohistochemistry, Paraffin embedded tissue.

  • Overlay histogram showing U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) 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, 0.1 μg/1x106) used under the same conditions. Unlabeled 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-87 MG cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • IHC-Fr image of Beta III Tubulin staining on zebrafish retina sections using ab52623 (1:100).

    The sections were fixed with paraformaldehyde and permeabilized using Triton-X. Antigen retrieval was performed using Tris-HCl. 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

  • 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 : PC-12 (Rat adrenal pheochromocytoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : 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.

  • Anti-beta III Tubulin antibody [EP1569Y] (ab52623) at 1/10000 dilution + HeLa (Human epithelial cell line from cervix adenocarcinoma) 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?

  • Flow Cytometry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling beta III Tubulin with ab52623 at 1/20 (red).

    Cells were fixed with 4% paraformaldehyde and permeabilized 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 - Unlabeled control, cells without incubation with primary and secondary antibodies.

References

This product has been referenced in:
  • Gao J  et al. In vitro investigation of the mechanism underlying the effect of ginsenoside on the proliferation and differentiation of neural stem cells subjected to oxygen-glucose deprivation/reperfusion. Int J Mol Med 41:353-363 (2018). Read more (PubMed: 29138802) »
  • Li HP  et al. Inactivation of the tight junction gene CLDN11 by aberrant hypermethylation modulates tubulins polymerization and promotes cell migration in nasopharyngeal carcinoma. J Exp Clin Cancer Res 37:102 (2018). Read more (PubMed: 29747653) »
See all 17 Publications for this product

Customer reviews and Q&As

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1-6 of 6 Abreviews

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

Abcam user community

Verified customer

Submitted 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

Mr. Julien Chevalier

Verified customer

Submitted 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

Mr. Julien Chevalier

Verified customer

Submitted 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

Mr. Julien Chevalier

Verified customer

Submitted 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

Ms. Babben Tinner

Verified customer

Submitted 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

Dr. Ryan Macdonald

Verified customer

Submitted Aug 22 2014

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

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