Overview

  • Product nameAnti-beta Tubulin antibody [EPR1330]
    See all beta Tubulin primary antibodies
  • Description
    Rabbit monoclonal [EPR1330] to beta Tubulin
  • Tested applicationsSuitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, African Green Monkey
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human beta Tubulin.

  • Positive control
    • MCF-7, Jurkat, Ramos, C6, Neuro-2a and HeLa cell lysates; Human fetal brain tissue lysate; Human brain, kidney and tonsil tissue, Mouse and Rat cerebral cortex tissue.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    A trial size is available to purchase for this antibody.

    An HRP variant is available as ab185057.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab108342 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 1/500.

For unpurified use at 1/100 - 1/250.

WB 1/1000 - 1/10000. Predicted molecular weight: 49 kDa.
IHC-P 1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol). 

For unpurified use at 1/250 - 1/500. 

Flow Cyt 1/20.

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

For unpurified use at 1/100 - 1/1000. 

Target

  • FunctionTubulin 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.
  • Tissue specificityUbiquitously expressed with highest levels in spleen, thymus and immature brain.
  • Involvement in diseaseCortical dysplasia, complex, with other brain malformations 6
    Skin creases, congenital symmetric circumferential, 1
  • Sequence similaritiesBelongs to the tubulin family.
  • DomainThe highly acidic C-terminal region may bind cations such as calcium.
  • Post-translational
    modifications
    Some glutamate residues at the C-terminus are polyglutamylated, resulting in polyglutamate chains on the gamma-carboxyl group (PubMed:26875866). Polyglutamylation plays a key role in microtubule severing by spastin (SPAST). SPAST preferentially recognizes and acts on microtubules decorated with short polyglutamate tails: severing activity by SPAST increases as the number of glutamates per tubulin rises from one to eight, but decreases beyond this glutamylation threshold (PubMed:26875866).
    Some glutamate residues at the C-terminus are 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). Both polyglutamylation and monoglycylation can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of monoglycylation is still unclear.
    Phosphorylated on Ser-172 by CDK1 during the cell cycle, from metaphase to telophase, but not in interphase. This phosphorylation inhibits tubulin incorporation into microtubules.
  • Cellular localizationCytoplasm, cytoskeleton.
  • Information by UniProt
  • Database links
  • Alternative names
    • Beta 4 tubulin antibody
    • Beta 5 tubulin antibody
    • beta Ib tubulin antibody
    • Beta1 tubulin antibody
    • Class I beta tubulin antibody
    • M40 antibody
    • MGC117247 antibody
    • MGC16435 antibody
    • OK/SW cl.56 antibody
    • OK/SWcl.56 antibody
    • TBB5_HUMAN antibody
    • TUBB 1 antibody
    • TUBB 2 antibody
    • TUBB 5 antibody
    • TUBB antibody
    • TUBB1 antibody
    • TUBB2 antibody
    • TUBB5 antibody
    • tubulin beta 1 chain antibody
    • Tubulin beta 2 chain antibody
    • tubulin beta 5 chain antibody
    • Tubulin beta chain antibody
    • Tubulin beta class I antibody
    • tubulin beta polypeptide antibody
    • Tubulin beta-5 chain antibody
    see all

Anti-beta Tubulin antibody [EPR1330] images

  • All lanes : Anti-beta Tubulin antibody [EPR1330] (ab108342) at 1/5000 dilution

    Lane 1 : Hela (human cervix adenocarcinoma) whole cell lysate
    Lane 2 : Human fetal brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size : 49 kDa
    Observed band size : 50 kDa (why is the actual band size different from the predicted?)
    Blocking/Diluting buffer 5% NFDM /TBST
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue sections labelling beta Tubulin with purified ab108342 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. Sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Immunocytochemistry/Immunofluorescence staining of HeLa cells labelling beta Tubulin with purified ab108342 at a working dilution of 1/500. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. DAPI was used as nuclear counterstain (bottom left hand panel). The cells were fixed in 4% Paraformaldehyde and permeabilized using 0.1% Triton X-100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, rabbit primary antibody was used followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120). For negative control 2, ab7291 (mouse anti-tubulin) was used followed by an Alexa Fluor® 488 goat anti-rabbit secondary (ab150077).

  • Overlay histogram showing 4% paraformaldehyde fixed Hela (human cervix adenocarcinoma) cells labelling beta Tubulin with purified ab108342 at dilution of 1/20. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG at dilution of 1/2000. A non-specific IgG antibody (rabbit monoclonal) was used as isotype control (black line). The blue line shows cells without incubation with primary antibody and secondary antibody. 

  • All lanes : Anti-beta Tubulin antibody [EPR1330] (ab108342) at 1/1000 dilution

    Lane 1 : Neuro-2a (mouse neuroblastoma) whole cell lysate
    Lane 2 : C6 (rat glioma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size : 49 kDa
    Observed band size : 50 kDa (why is the actual band size different from the predicted?)
    Blocking/Diluting buffer 5% NFDM /TBST
  • Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue sections labelling beta Tubulin with purified ab108342 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. Sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • All lanes : Anti-beta Tubulin antibody [EPR1330] (ab108342) at 1/1000 dilution

    Lane 1 : MCF-7 cell lysates
    Lane 2 : Jurkat cell lysates
    Lane 3 : Ramos cell lysates
    Lane 4 : HeLa cell lysates

    Secondary
    HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size : 49 kDa
  • Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue sections labelling beta Tubulin with purified ab108342 at dilution of 1/50. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. Sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Immunofluorescent staining of HeLa cells using ab108342 at 1/100 dilution.
  • ab108342 staining beta Tubulin in human HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Samples were incubated with primary antibody (1/200 in PBS) for 1 hour at 22°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Counterstained with DAPI.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab108342 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab108342, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) 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 HeLa fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.
  • ab108342 at 1/250 dilution staining beta Tubulin in Human brain tissue by Immunohistochemistry Formalin-fixed, Paraffin-embedded tissue.
  • ab108342 at 1/250 dilution staining beta Tubulin in Human tonsil tissue by Immunohistochemistry Formalin-fixed, Paraffin-embedded tissue.
  • ab108342 showing positive staining in Normal kidney tissue.

  • ab108342 showing positive staining in Breast carcinoma tissue.

References for Anti-beta Tubulin antibody [EPR1330] (ab108342)

This product has been referenced in:
  • Giacomello E  et al. Deletion of small ankyrin 1 (sAnk1) isoforms results in structural and functional alterations in aging skeletal muscle fibers. Am J Physiol Cell Physiol 308:C123-38 (2015). Mouse . Read more (PubMed: 25354526) »
  • Wang JF  et al. The molecular mechanisms of Tanshinone IIA on the apoptosis and arrest of human esophageal carcinoma cells. Biomed Res Int 2014:582730 (2014). WB ; Human . Read more (PubMed: 24829906) »

See all 2 Publications for this product

Product Wall

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (HEK293 cells)
Specification HEK293 cells
Treatment TGF-beta, 24 hours, 50 pM or untreated
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted May 07 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (NMuMG cells)
Specification NMuMG cells
Treatment TGF-beta, 24 hours, 50 pM or untreated
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted May 07 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample African Green Monkey Cell lysate - whole cell (COS-1 cells)
Specification COS-1 cells
Treatment TGF-beta, 24 hours, 50 pM or untreated
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted May 07 2014

Application Western blot
Loading amount 62.5 µg
Gel Running Conditions Reduced Denaturing (12.5% acrylamide)
Sample Pig Tissue lysate - other (Muscle sarcoplasmic extract)
Specification Muscle sarcoplasmic extract
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. Shannon Cruzen

Verified customer

Submitted Feb 12 2014

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Permeabilization Yes - 0.5% Triton X100 in PBS
Fixative Paraformaldehyde
Username

Dr. Kirk McManus

Verified customer

Submitted Jan 22 2014

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