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Tags & Cell Markers Subcellular Markers Cytoskeleton Microtubules
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-beta III Tubulin antibody [EP1569Y] (ab52623)

  • Datasheet
  • SDS
Reviews (6)Q&A (1)References (28)

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Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
  • Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
  • Multiplex immunohistochemistry - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
  • Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
  • Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
  • Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
  • Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
  • Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
  • Flow Cytometry - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
  • Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
  • Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
  • Flow Cytometry - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
  • Anti-beta III Tubulin antibody [EP1569Y] (ab52623)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1569Y] to beta III Tubulin
  • Suitable for: mIHC, ICC/IF, WB, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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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 & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Mouse
    Rat
    Human
    IHC-P
    Human
    mIHC
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide within Human beta III Tubulin aa 400 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: Q13509

  • Positive control

    • ICC/IF: PC-12, HAP1 and HeLa cells. WB: HAP1 cell lysate. HeLa, HCT116, PC-12 and HEK-293 whole cell lysate. Mouse and rat brain and spinal cord tissue lysate. IHC-P: Human breast carcinoma tissue. mIHC: Human cerebellum tissue Flow Cyt: U-87 MG and HeLa cells. IHC-Fr: Zebrafish retina sections.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EP1569Y
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Cytoskeleton
    • Microtubules
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microtubules
    • Tubulin

Associated products

  • Alternative Versions

    • HRP Anti-beta III Tubulin antibody [EP1569Y] (ab190574)
    • Alexa Fluor® 647 Anti-beta III Tubulin antibody [EP1569Y] (ab190575)
    • Biotin Anti-beta III Tubulin antibody [EP1569Y] (ab195903)
    • Alexa Fluor® 594 Anti-beta III Tubulin antibody [EP1569Y] (ab201740)
    • Alexa Fluor® 555 Anti-beta III Tubulin antibody [EP1569Y] - Neuronal Marker (ab202519)
    • Anti-beta III Tubulin antibody [EP1569Y] - BSA and Azide free (ab221935)
    • APC Anti-beta III Tubulin antibody [EP1569Y] (ab224977)
    • FITC Anti-beta III Tubulin antibody [EP1569Y] (ab224978)
    • PE Anti-beta III Tubulin antibody [EP1569Y] (ab224979)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human TUBB3 (beta III Tubulin) knockout HeLa cell line (ab255358)
    • Human TUBB3 (beta III Tubulin) knockout HCT116 cell line (ab266900)
  • KO cell lysates

    • Human TUBB3 (beta III Tubulin) knockout HCT116 cell lysate (ab257070)
    • Human TUBB3 (beta III Tubulin) knockout HeLa cell lysate (ab263857)
  • KO cell pellets

    • Human TUBB3 (beta III Tubulin) knockout HeLa cell pellet (ab278939)
  • Recombinant Protein

    • Recombinant Human beta III Tubulin protein (ab140581)
  • Related Products

    • ioGlutamatergic Neurons - Human iPSC-Derived Glutamatergic Neurons (ab259259)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Mouse
Rat
Human
IHC-P
Human
mIHC
Human
WB
Mouse
Rat
Human
All applications
Zebrafish
Application Abreviews Notes
mIHC
1/1000.

Perform Sodium citrate antigen retrieval (pH 6.0) between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

 

ICC/IF (1)
1/500.
WB (3)
1/1000 - 1/10000. Detects a band of approximately 52 kDa.
Flow Cyt
1/10 - 1/1000.

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

IHC-P (1)
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
mIHC
1/1000.

Perform Sodium citrate antigen retrieval (pH 6.0) between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

 

ICC/IF
1/500.
WB
1/1000 - 1/10000. Detects a band of approximately 52 kDa.
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. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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.
  • Target information above from: UniProt accession Q13509 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10381 Human
    • Entrez Gene: 22152 Mouse
    • Entrez Gene: 246118 Rat
    • Omim: 602661 Human
    • SwissProt: Q13509 Human
    • SwissProt: Q9ERD7 Mouse
    • SwissProt: Q4QRB4 Rat
    • Unigene: 511743 Human
    • Unigene: 40068 Mouse
    • Unigene: 43958 Rat
    see all
  • Alternative names

    • beta 3 tubulin antibody
    • beta 4 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
    • tuj 1 antibody
    • tuj1 antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)

    Immunofluorescence staining of beta III Tubulin using ab52623 in ioNEURONS/glut cells (Human iPSC-Derived Glutamatergic Neurons, ab259259), which were differentiated for 11 days post induction.

    The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 mins 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 overnight at +4°C with ab52623 at 0.1 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

  • Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    All lanes : Anti-beta III Tubulin antibody [EP1569Y] (ab52623) at 1/1000 dilution

    Lane 1 : Wild-type HCT116 cell lysate
    Lane 2 : TUBB3 knockout HCT116 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.


    Lanes 1- 2: Merged signal (red and green). Green - ab52623 observed at 52 kDa. Red - loading control ab8245 observed at 37 kDa.
    ab52623 Anti-beta III Tubulin antibody [EP1569Y] was shown to specifically react with beta III Tubulin in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266900 (knockout cell lysate ab257070) was used. Wild-type and beta III Tubulin knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab52623 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Multiplex immunohistochemistry - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Multiplex immunohistochemistry - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)

    Fluorescence multiplex immunohistochemical analysis of human cerebellum tissue (formalin-fixed paraffin-embedded section).

    Merged staining of Neu-N (ab177487; yellow; Opal™570), anti-beta III Tubulin (ab52623; red; Opal™690) and anti-GFAP (ab68428; green; Opal™520).

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ kit.

    The section was incubated in three rounds of staining with ab177487 (1/1000 dilution), ab52623 (1/200 dilution) and ab68428 (1/250 dilution); each using a separate fluorescent tyramide signal amplification system.

    Sodium citrate antigen retrieval (pH 6.0) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

    DAPI (blue) was used as a nuclear counter stain.

  • Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)

    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).

  • Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    All lanes : Anti-beta III Tubulin antibody [EP1569Y] (ab52623) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : TUBB3 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Observed band size: 50 kDa why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab52623 observed at 50 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab52623 was shown to react with beta III tubulin in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255358 (knockout cell lysate ab263857) was used. Wild-type HeLa and TUBB3 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab52623 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    All lanes : Anti-beta III Tubulin antibody [EP1569Y] (ab52623) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : Beta III Tubulin knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : HEK293 whole cell lysate

    Lysates/proteins at 20 µg per lane.


    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.

  • Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)Eng et al PLoS One. 2017 May 17;12(5):e0177834. doi: 10.1371/journal.pone.0177834. eCollection 2017. Fig 5. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)

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

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Flow Cytometry - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)

    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.

  • Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    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.

  • Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Western blot - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    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 - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Flow Cytometry - Anti-beta III Tubulin antibody [EP1569Y] (ab52623)

    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.

  • Anti-beta III Tubulin antibody [EP1569Y] (ab52623)
    Anti-beta III Tubulin antibody [EP1569Y] (ab52623)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (28)

Publishing research using ab52623? Please let us know so that we can cite the reference in this datasheet.

ab52623 has been referenced in 28 publications.

  • Philpott JM  et al. Casein kinase 1 dynamics underlie substrate selectivity and the PER2 circadian phosphoswitch. Elife 9:N/A (2020). PubMed: 32043967
  • Zeng X  et al. Innervation of thermogenic adipose tissue via a calsyntenin 3ß-S100b axis. Nature 569:229-235 (2019). PubMed: 31043739
  • Doering L  et al. Regulation of somatostatin expression by vitamin D3 and valproic acid in human adipose-derived mesenchymal stem cells. Stem Cell Res Ther 10:240 (2019). PubMed: 31387633
  • Kobayashi Y  et al. Selectively high efficacy of eribulin against high-grade invasive recurrent and/or metastatic squamous cell carcinoma of the head and neck. Oncol Lett 17:5064-5072 (2019). PubMed: 31186718
  • Springer NL  et al. Obesity-Associated Extracellular Matrix Remodeling Promotes a Macrophage Phenotype Similar to Tumor-Associated Macrophages. Am J Pathol 189:2019-2035 (2019). PubMed: 31323189
View all Publications for this product

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-beta III Tubulin antibody [EP1569Y]

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Oct 27 2017

Western blot abreview for Anti-beta III Tubulin antibody [EP1569Y]

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Mr. Julien Chevalier

Verified customer

Submitted Sep 30 2015

Western blot abreview for Anti-beta III Tubulin antibody [EP1569Y]

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Mr. Julien Chevalier

Verified customer

Submitted Sep 30 2015

Western blot abreview for Anti-beta III Tubulin antibody [EP1569Y]

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Mr. Julien Chevalier

Verified customer

Submitted Sep 30 2015

Immunocytochemistry/ Immunofluorescence abreview for Anti-beta III Tubulin antibody [EP1569Y]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Cryopreserved embryonic cortical neurons (QBMcells)
Specification
Cryopreserved embryonic cortical neurons (QBMcells
Permeabilization
No
Fixative
paraformaldehyde with picric acid
Read More

Ms. Babben Tinner

Verified customer

Submitted Feb 26 2015

Immunohistochemistry (Frozen sections) abreview for Anti-beta III Tubulin antibody [EP1569Y]

Good
Abreviews
Abreviews
abreview image
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
Read More

Dr. Ryan Macdonald

Verified customer

Submitted Aug 22 2014

Question

Hi, I’d appreciate some help searching your products page on your website!  I’m trying to find primary antibodies against the C-terminal ends of alpha and beta tubulin for immunogold electron microscopy, but your search engine gives me 180 results for anti-tubulin antibodies but no means of sorting them with respect to their epitope.  I’d rather not have to open each webpage for all 180 hits to find the antibody I need, is there an easier way?   This work will be done in mouse, rat and human tissues, so I need the primaries not raised from these species.   Many thanks,      

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Abcam community

Verified customer

Asked on Sep 15 2011

Answer

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 consider trying the following rabbit monoclonal antibody which is tested in mouse, however I am sorry this is not tested and guaranteed for EM. ab52623 Rabbit mono Anti-beta Tubulin antibody [EP1569Y] Applications Flow Cyt, ICC, IHC-P, IP, WB Reacts with: Human, Mouse, Rat Immunogen propriatory, but it is right on the C-terminus and about 15 residues long. https://www.abcam.com/beta-III-Tubulin-antibody-EP1569Y-ab52623.html https://www.abcam.com/beta-III-Tubulin-antibody-EP1569Y-ab52623.html. For alpha tubulin, we have the following antibody tested in EM and mouse, rat and human but rasied in mouse. I am sorry the epitope has not been mapped: ab80779 Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free Mouse monoclonal Applications: EM, Flow Cyt, ICC/IF, IHC-P, IP, WB Reacts with: Human, Mouse, Rat, Chicken, Cow, Gerbil, Guinea pig, Pig, Xenopus laevis https://www.abcam.com/alpha-Tubulin-antibody-DM1A-ab80779.html https://www.abcam.com/alpha-Tubulin-antibody-DM1A-ab80779.html. I understand you concerns that mouse on mouse can sometimes give non specific binding. However, this has been tested succesfully in mouse, and we are able to provide our guarantee for the results from mouse and EM. Otherwise, it could be preferable to try an antibody raised in a different species. The following rabbit polyclonal is tested in mouse, rat and human and is raised against a peptide from the C terminus. However, I am sorry it is not tested and guaranteed in EM: ab15246 Anti-alpha Tubulin antibody Rabbit polyclonal Applications: ICC/IF, IHC-Fr, IHC-P, IP, WB Reacts with: Fruit fly (Drosophila melanogaster), Human, Mouse, Pig, Rat, Zebrafish Immunogen: ALEKDYEEVGVDSVEGEGEEEGEEY C terminal amino acids 426-450 of Human alpha Tubulin https://www.abcam.com/alpha-Tubulin-antibody-Microtubule-Marker-ab15246.html https://www.abcam.com/alpha-Tubulin-antibody-Microtubule-Marker-ab15246.html. With regards to the search facility on the website, I understand it is not possible to search by immunogen or region. This is partly because although we produce many antibodies in house and can provide the immunogen sequence, we do source many externally and sometimes the immunogen information is propriatory (commercially sensitive). We can usually provide this information on individual customer request. Thank you for your understanding. I hope this will be helpful to you. Should you have any further questions, please do not hesitate to contact us.

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Abcam Scientific Support

Answered on Sep 15 2011

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