Overview

  • Product name
    Anti-alpha Tubulin (acetyl K40) antibody [EPR16772]
    See all alpha Tubulin primary antibodies
  • Description
    Rabbit monoclonal [EPR16772] to alpha Tubulin (acetyl K40)
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, Flow Cyt, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human alpha Tubulin aa 1-100 (acetyl K40). The exact sequence is proprietary.
    Database link: P68363

  • Positive control
    • WB: HeLa, C6 and NIH/3T3 whole cell lysates (treated with 500 ng/ml Trichostatin A for 4 hours); Mouse brain, kidney and spleen lysates; Rat brain and heart lysates; Human fetal heart and fetal kidney lysates. IHC-P: Human and Mouse cerebral cortex tissue; rat cerebellum tissue. IF: HeLa cells treated with 50 ug/ml Trichostatin A for 4 hours. Flow: HeLa cells treated with 500ng/ml Trichostatin A for 4 hours. IP: HeLa treated with 500 ng/ml Trichostatin A for 4 hours.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab179484 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
WB 1/2000. Detects a band of approximately 52 kDa (predicted molecular weight: 50 kDa).
ICC/IF 1/500.
Flow Cyt 1/240.

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

 

IP 1/70.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 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.
  • Sequence similarities
    Belongs 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 localization
    Cytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links
  • Alternative names
    • Alpha-tubulin 1 antibody
    • ALS22 antibody
    • B ALPHA 1 antibody
    • bA408E5.3 antibody
    • H2 ALPHA antibody
    • Hum a tub1 antibody
    • Hum a tub2 antibody
    • LIS3 antibody
    • MGC171407 antibody
    • MGC55332 antibody
    • TBA4A_HUMAN antibody
    • Testis-specific alpha-tubulin antibody
    • TUBA1 antibody
    • TUBA1A antibody
    • tuba1l antibody
    • Tuba4a antibody
    • Tubulin alpha 1 chain antibody
    • Tubulin alpha antibody
    • Tubulin alpha-1 chain antibody
    • tubulin alpha-1B chain antibody
    • Tubulin alpha-4A chain antibody
    • Tubulin H2-alpha antibody
    • Tubulin, alpha 1 (testis specific) antibody
    • tubulin, alpha 1, like antibody
    • Tubulin, alpha 4a antibody
    • Tubulin, alpha, testis-specific antibody
    • Tubulin, alpha-1 antibody
    see all

Images

  • All lanes : Anti-alpha Tubulin (acetyl K40) antibody [EPR16772] (ab179484) at 1/20000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate treated with 500 ng/ml Trichostatin A for 4 hours
    Lane 2 : Untreated HeLa whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 50 kDa
    Observed band size: 52 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Ab179484 staining alpha Tubulin in HFF-1 (Human skin fibroblast) cell line by ICC/IF (Immunocytochemistry/Immunofluorescence). The cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody at 1:20000 dilution. An AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as a secondary antibody at 1:1000 dilution. An Anti-Alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594), ab195889 was used as a counterstain at 1:200 dilution. DAPI was used as a nuclear counterstain. Confocal image showing cilia (arrows) staining in HFF-1 cells treated with starvation for 48 hours.

  • Ab179484 staining alpha Tubulin in NIH/3T3 (mouse embryonic fibroblast) cell line by ICC/IF (Immunocytochemistry/Immunofluorescence).The cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody at 1:20000 dilution. An AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as a secondary antibody at 1:1000 dilution. An Anti-Alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594), ab195889 was used as a counterstain at 1:200 dilution. DAPI was used as a nuclear counterstain. Confocal image showing cilia (arrows) staining in NIH/3T3 cells treated with starvation for 48 hours.

  • Ab179484 staining alpha Tubulin in NIH/3T3 (mouse embryonic fibroblast) cell line by ICC/IF (Immunocytochemistry/Immunofluorescence). The cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody at 1:20000 dilution. An AlexaFluor®488 Goat anti-Rabbit (ab150077) was used as a secondary antibody at 1:1000 dilution. An Anti-Alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594), ab195889 was used as a counterstain at 1:200 dilution. DAPI was used as a nuclear counterstain. Confocal image showing midbody (arrows) staining in NIH/3T3 cells treated with starvation for 48 hours.

  • ab179484 stained in Hela cells. Untreated and Trichostatin A treated (50ug/ml, 4 hours) cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab179484 at 1/500 dilution overnight at +4°C. The secondary antibody was ab150177 used at 1 ug/ml for 1hour at room temperature (colored green). DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • All lanes : Anti-alpha Tubulin (acetyl K40) antibody [EPR16772] (ab179484) at 1/20000 dilution

    Lane 1 : C6 (Rat glial tumor cells) whole cell lysate treated with 500 ng/ml Trichostatin A for 4 hours
    Lane 2 : Untreated C6 whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 50 kDa
    Observed band size: 52 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-alpha Tubulin (acetyl K40) antibody [EPR16772] (ab179484) at 1/20000 dilution

    Lane 1 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate treated with 500 ng/ml Trichostatin A for 4 hours
    Lane 2 : Untreated NIH/3T3 whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lane 1 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
    Lane 2 : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 50 kDa
    Additional bands at: 52 kDa. We are unsure as to the identity of these extra bands.



    Blocking/dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-alpha Tubulin (acetyl K40) antibody [EPR16772] (ab179484) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat heart lysate
    Lane 6 : Human fetal heart lysate
    Lane 7 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 50 kDa
    Observed band size: 52 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling alpha Tubulin (acetyl K40) with ab179484 at 1/1000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic staining is observed on Purkinje cells of cerebellum. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling alpha Tubulin (acetyl K40) with ab179484 at 1/1000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic staining is observed on neuron cells of Human brain tissue. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling alpha Tubulin (acetyl K40) with ab179484 at 1/1000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic staining is observed on neuron cells of Mouse cerebral cortex tissue. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • Flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells treated with 500 ng/ml Trichostatin A for 4 hours labeling alpha Tubulin (acetyl K40) with ab179484 at 1/240 dilution (red line). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.  ab179484 preincubated with 1mg/ml acetyl Alpha tubulin (acetyl K40) peptide (green) or non-acetyl Alpha tubulin (acetyl K40) peptide (orange). The isotype control was Rabbit monoclonal IgG (black) and the unlabelled contol was cells without incubation with primary antibody and secondary antibody (blue).

  • Alpha Tubulin was immunoprecipitated from 1mg of HeLa cells (Human epithelial cells from cervix adenocarcinoma) treated with 500 ng/ml Trichostatin A for 4 hours with ab179484 at 1/70 dilution. Western blot was performed from 10 µg of the immunoprecipitate using ab179484 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Left lane: Hela whole cell extract. Right lane: PBS instead of Hela whole cell extract.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

References

This product has been referenced in:
  • Dong J  et al. A novel HDAC6 inhibitor exerts an anti-cancer effect by triggering cell cycle arrest and apoptosis in gastric cancer. Eur J Pharmacol 828:67-79 (2018). WB . Read more (PubMed: 29563065) »
  • Chen S  et al. Developmental abnormalities in supporting cell phalangeal processes and cytoskeleton in the Gjb2 knockdown mouse model. Dis Model Mech 11:N/A (2018). IHC-P, IF . Read more (PubMed: 29361521) »
See all 6 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Bos taurus Tissue sections (oviduct)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA (pH 9.0)
Permeabilization
No
Specification
oviduct
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 04 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Antigen retrieval step
None
Sample
Mouse Tissue sections (adult mouse lung)
Specification
adult mouse lung
Permeabilization
Yes - 0.5% Triton X-100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 31 2014

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

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