Overview

  • Product nameAnti-alpha Tubulin antibody
    See all alpha Tubulin primary antibodies
  • Description
    Rabbit polyclonal to alpha Tubulin
  • Tested applicationsICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Cow, Human, Fruit fly (Drosophila melanogaster), Indian Muntjac, African Green Monkey, Chinese Hamster
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 400 to the C-terminus of Human alpha Tubulin.

  • Positive control
    • This antibody gave a positive signal within WB in the following whole cell lysates: HeLa; HEK293; HepG2; Caco2; NIH3T3; PC12. HeLa cells - IF. ICC/IF - Caco-2, NIH3T3 and SV40

Properties

Applications

Our Abpromise guarantee covers the use of ab18251 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
IF Use a concentration of 1 - 5 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P 1/2500. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
WB Use a concentration of 0.5 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).Can be blocked with Human alpha Tubulin peptide (ab18648).

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.
  • Sequence similaritiesBelongs to the tubulin family.
  • 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.
    Acetylation of alpha chains at Lys-40 stabilizes microtubules and affects affinity and processivity of microtubule motors. This modification has a role in multiple cellular functions, ranging from cell motility, cell cycle progression or cell differentiation to intracellular trafficking and signaling.
  • Cellular localizationCytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links
  • Alternative names
    • Alpha tubulin ubiquitous antibody
    • Alpha-tubulin 1 antibody
    • B ALPHA 1 antibody
    • bA408E5.3 antibody
    • H2 ALPHA antibody
    • Hum a tub1 antibody
    • Hum a tub2 antibody
    • LIS3 antibody
    • TBA4A_HUMAN antibody
    • Testis-specific alpha-tubulin antibody
    • TUBA1 antibody
    • TUBA1A antibody
    • Tuba4a antibody
    • Tubulin alpha 1 chain antibody
    • Tubulin alpha antibody
    • Tubulin alpha ubiquitous chain antibody
    • Tubulin alpha-1 chain antibody
    • Tubulin alpha-4A chain antibody
    • Tubulin H2-alpha antibody
    • Tubulin K alpha 1 antibody
    • Tubulin, alpha 1 (testis specific) antibody
    • Tubulin, alpha 4a antibody
    • Tubulin, alpha, testis-specific antibody
    • Tubulin, alpha-1 antibody
    see all

Anti-alpha Tubulin antibody images

  • ab18251 staining alpha-Tubulin in SV40 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 ab18251 at 1μl/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 antibody 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.

  • ab18251 staining alpha-Tubulin in NIH3T3 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 ab18251 at 1μl/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 antibody 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.

  • ab18251 staining alpha-Tubulin in Caco-2 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 ab18251 at 5μl/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 antibody 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-alpha Tubulin antibody (ab18251) at 0.5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
    Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 6 : 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

    Predicted band size : 50 kDa
    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 ab18251 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.

  • ab18251 staining alpha Tubulin in the COS7 fibroblast cell line from African Green Monkey  by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methano and blocked with 5% BSA for 60 minutes at 21°C. Samples were incubated with primary antibody (1/500) for 17 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal(1/400) was used as the secondary antibody.

    See Abreview

  • ICC/IF image of ab18251 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab18251, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue).
  • All lanes : Anti-alpha Tubulin antibody (ab18251) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 50 kDa
    Observed band size : 50 kDa


    Exposure time : 1 minute

    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 2% Bovine Serum Albumin before being incubated with ab18251 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

  • All lanes : Anti-alpha Tubulin antibody (ab18251) at 0.5 µg/ml

    Lane 1 : HeLa lysate
    Lane 2 : A431 lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Alexa Flour Goat polyclonal to Rabbit IgG (700) at 1/10000 dilution

    Predicted band size : 50 kDa
    Observed band size : 50 kDa
    Additional bands at : 30 kDa (possible cross reactivity).

    ab18251 detects a strong band at 50 kDa corresponding to alpha tubulin. Cross-reactivity is also seen with other lower molecular weight bands. This may be reduced by using the antibody at a lower working concentration.

  • ab18251 at a 1/8000 dilution staining human HeLa cells by immunocytochemistry. The cells were paraformaldehyde fixed and incubated with the antibody for 30 minutes. The secondary antibody was a Cy3 conjugated Goat Anti-Rabbit IgG (H+L). The image shows staining of an interphase IM cell.

    This image is courtesy of an Abreview by Kirk McManus submitted on 27 February 2006.

    See Abreview

  • Image courtesy of Human Protein Atlas

    ab18251 staining alpha Tubulin. Paraffin embedded human appendix tissue was incubated with ab18251 (1/2500 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab18251 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

    Further results for this antibody can be found at www.proteinatlas.org

  • ab18251 staining alpha Tubulin in HeLa cells by Immunocytochemistry/ Immunofluorescence.
    ab18251 staining alpha Tubulin labeled red.
    ab20740 staining Lamin A labeled green.
    Hoechst 33258 staining DNA blue.

References for Anti-alpha Tubulin antibody (ab18251)

This product has been referenced in:
  • Kim S & Rhee K Importance of the CEP215-pericentrin interaction for centrosome maturation during mitosis. PLoS One 9:e87016 (2014). ICC/IF ; Human . Read more (PubMed: 24466316) »
  • Álvarez-Quilón A  et al. ATM specifically mediates repair of double-strand breaks with blocked DNA ends. Nat Commun 5:3347 (2014). ICC/IF ; Mouse . Read more (PubMed: 24572510) »

See all 31 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: HIER Tris/HCL pH 9.0
Sample Cow Tissue sections (Bovine Liver)
Specification Bovine Liver
Permeabilization Yes - 0.2% Triton X-100 and PBS
Fixative Formaldehyde
Username

Dr. Steffen Rickelt

Verified customer

Submitted Aug 08 2014

Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Human Cell lysate - whole cell (Hela)
Specification Hela
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Dec 12 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Sample Human Tissue sections (heart)
Specification heart
Permeabilization Yes - 0.3% trinton X-100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 10 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (hippocampal neurons in culture)
Specification hippocampal neurons in culture
Fixative Methanol
Permeabilization Yes - Triton X-100
Blocking step (agent) for 45 minute(s) · Concentration: 0.22% · Temperature: 20°C
Username

Dr. Christophe Leterrier

Verified customer

Submitted May 03 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample African Green Monkey Cell (Cos-7 cell)
Specification Cos-7 cell
Fixative Methanol
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Mar 26 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample African Green Monkey Cell lysate - whole cell (Caco-2 cell, Cos-7 cell)
Loading amount 3 µg
Specification Caco-2 cell, Cos-7 cell
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Mar 26 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Caco-2 cell)
Specification Caco-2 cell
Fixative Methanol
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Mar 26 2013

Thank you for contacting us. I want to apologize for the delay as we arranged to add each peptide to our catalog.

The blocking peptides which you have requested are now available and can be located at the following links:


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Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293)
Loading amount 20 µg
Specification HEK293
Gel Running Conditions Non-reduced Denaturing (10% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted May 17 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Fruit fly (Drosophila melanogaster) Cell (wing)
Specification wing
Fixative Paraformaldehyde
Permeabilization Yes - 0.1 % Triton X 100
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2µg/mL · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Oct 18 2011

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