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  1. Link

    beta-iii-tubulin-antibody-5g8-ab231084.pdf

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Signal Transduction Cytoskeleton / ECM Cytoskeleton Microtubules Tubulin
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Validated using a knockout cell line

Anti-beta III Tubulin antibody [5G8] (ab231084)

  • Datasheet
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [5G8] (ab231084)
  • Western blot - Anti-beta III Tubulin antibody [5G8] (ab231084)
  • Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [5G8] (ab231084)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [5G8] (ab231084)

Key features and details

  • Mouse monoclonal [5G8] to beta III Tubulin
  • Suitable for: ICC/IF, IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG1

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Protein
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Recombinant Human beta III Tubulin protein (ab140581)
Secondary
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Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
Knockout
Product image
Human TUBB3 (beta III Tubulin) knockout HeLa cell line (ab255358)

View more associated products

Overview

  • Product name

    Anti-beta III Tubulin antibody [5G8]
    See all beta III Tubulin primary antibodies
  • Description

    Mouse monoclonal [5G8] to beta III Tubulin
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human beta III Tubulin aa 400 to the C-terminus.
    Database link: Q13509

  • Epitope

    The epitope recognized by 5G8 is EAQGPK
  • Positive control

    • ICC/IF: HAP1 WT cells. IHC-P: FFPE human cerebellum and rat cerebellum tissue sections. WB: HAP1, Neuro2a and PC12 cell lysates
  • General notes

    This antibody clone is manufactured by Abcam. If you require a different buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    5G8
  • Isotype

    IgG1
  • Light chain type

    kappa
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microtubules
    • Tubulin

Associated products

  • Alternative Versions

    • Anti-beta III Tubulin antibody [5G8] - BSA and Azide free (ab264113)
  • Recombinant Protein

    • Recombinant Human beta III Tubulin protein (ab140581)

Applications

Our Abpromise guarantee covers the use of ab231084 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
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 50 kDa).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [5G8] (ab231084)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [5G8] (ab231084)

    IHC image of beta III Tubulin staining in a section of formalin-fixed paraffin-embedded normal human cerebellum 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 20mins. The section was then incubated with ab231084, 1ug/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. The inset secondary-only control image is taken from an identical assay without primary antibody.

    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.

  • Western blot - Anti-beta III Tubulin antibody [5G8] (ab231084)
    Western blot - Anti-beta III Tubulin antibody [5G8] (ab231084)
    All lanes :

    Lane 1 : HAP1 whole cell lysate
    Lane 2 : HAP1 TUBB3 knockout whole cell lysate
    Lane 3 : Neuro2a whole cell lysate
    Lane 4 : PC12 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 50 kDa
    Observed band size: 55 kDa
    why is the actual band size different from the predicted?



    ab231084 was shown to specifically react with beta III Tubulin (TUBB3) in wild type HAP1 cells. No band was observed when beta III Tubulin (TUBB3) knockout samples were used. Wild-type and beta III Tubulin (TUBB3) knockout samples were subjected to SDS-PAGE. ab231084 and ab181602 (Rabbit anti GAPDH) were incubated overnight at 4°C at 1ug/ml and 1/10000 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 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [5G8] (ab231084)
    Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [5G8] (ab231084)

    ab231084 staining beta III tubulin (shown in green) in wild-type HAP1 cells (top panel) and TUBB3 knockout HAP1 cells (bottom panel).

    The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 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 ab231084 at 1 μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labeled with DAPI (shown in blue).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [5G8] (ab231084)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [5G8] (ab231084)

    IHC image of beta III Tubulin staining in a section of formalin-fixed paraffin-embedded normal rat cerebellum 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 20mins. The section was then incubated with ab231084, 1ug/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. The inset secondary-only control image is taken from an identical assay without primary antibody.

    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.

     

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

    • Datasheet
  • References (1)

    Publishing research using ab231084? Please let us know so that we can cite the reference in this datasheet.

    ab231084 has been referenced in 1 publication.

    • Spano AJ & Frankfurter A Characterization of anti-beta-tubulin antibodies. Methods Cell Biol 95:33-46 (2010). PubMed: 20466128

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