Anti-alpha Tubulin (acetyl K40) antibody [6-11B-1] (ab24610)

Overview

  • Product name
    Anti-alpha Tubulin (acetyl K40) antibody [6-11B-1]
    See all alpha Tubulin primary antibodies
  • Description
    Mouse monoclonal [6-11B-1] to alpha Tubulin (acetyl K40)
  • Specificity
    ab24610 detects acetylated alpha tubulin.
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Human, Monkey, Sea urchin
    Predicted to work with: Cow
  • Immunogen

    Tissue, cells or virus corresponding to alpha Tubulin.

  • Epitope
    The antibody recognizes an epitope located on the a3 isoform of Chlamydomonas axonemal a-tubulin, within four residues of Lys40 when this amino acid is acetylated.
  • Positive control
    • In Western Blot, this antibody gave a positive signal in mouse brain tissue lysate and in the following whole cell lysates: HeLa; NIH3T3; PC12.
  • General notes

    Production of this antibody has been changed on 8th April 2016. This antibody is now purified from tissue culture supernatant. This shouldn’t affect the use of this antibody but if you have any issues, please contact our Scientific Support team.

    This antibody binds to primary cilia, centrioles, mitotic spindles, midbodies and to subsets of cytoplasmic microtubules in 3T3 and HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab24610 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
Flow Cyt Use 1µl for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

WB Use a concentration of 0.03 - 0.06 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

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 [6-11B-1] (ab24610) at 5 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 4 : Brain (Mouse) Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 55 kDa
    Observed band size : 55 kDa
    Additional bands at : 140 kDa,25 kDa,35 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 150 seconds
  • ab24610 staining Acetylated alpha Tubulin in monkey kidney cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 3% PFA + 0.1% GA and blocked with 3% BSA + 0.5% Triton X-100 for 45 minutes at 25°C. Samples were incubated with primary antibody (1/100 in 3% BSA + 0.5% Triton X-100) for 1 hour at 21°C. An Alexa Fluor® 647-conjugated donkey anti-rabbit IgG polyclonal (2 µg/ml) was used as the secondary antibody.

    See Abreview

  • ab24610 at 1/100 dilution staining acetylated alpha tubulinin in prostate carcinoma by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). Sections were paraformaldehyde fixed, permeabilized in Triton X-100 prior to blocking in 1% serum for 1 hour at 27°C and then incubated with ab24610 for 12 hours at 4°C. Alexa Fluor® 546 donkey polyclonal to mouse Ig, diluted 1/500, was used as the secondary antibody.

    See Abreview

  • ICC/IF image of ab24610 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab24610, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT™ 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). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • ICC/IF image of ab24610 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab24610, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing HeLa cells stained with ab24610 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween 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 (ab24610, 1µg/1x106 cells) for 30 min at 22ºC. (This data was generated from a purified version of the antibody. Some lots are produced as ascites fluid. We suggest 1µl/1x106 cells for ascites preparations). The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:
  • Farrugia AJ & Calvo F Cdc42 regulates Cdc42EP3 function in cancer-associated fibroblasts. Small GTPases 8:49-57 (2017). ICC/IF . Read more (PubMed: 27248291) »
  • Patschan D  et al. Endothelial Colony Forming Cells (ECFCs) in murine AKI - implications for future cell-based therapies. BMC Nephrol 18:53 (2017). IHC-P ; Mouse . Read more (PubMed: 28166726) »

See all 85 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (immortalised bone marrow-derived macrophage)
Permeabilization
Yes - 0.1% Triton X-100
Specification
immortalised bone marrow-derived macrophage
Blocking step
No blocking step used for 1 hour(s) and 0 minute(s) · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Dr. James Harris

Verified customer

Submitted Jun 02 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (Swine Testicle Cells (ST))
Permeabilization
Yes - 0.1% Triton X-100
Specification
Swine Testicle Cells (ST)
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 28 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Fibroblasts)
Permeabilization
Yes - PBS1X, 0.2% triton
Specification
Fibroblasts
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 29 2016

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
3% BSA and 0.5% TX100 as blocking agent for 45 minute(s) · Concentration: 3% · Temperature: 25°C
Sample
Monkey Cell (Kidney)
Specification
Kidney
Permeabilization
No
Fixative
3% PFA and 0.1% GA
Username

Dr. Aaron Halpen

Verified customer

Submitted Sep 29 2014

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cultured Cells (Mouse Glioblastoma (GL261))
Permeabilization
Yes - 0.3% TritonX-100 in 1X TBS
Specification
Mouse Glioblastoma (GL261)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 28°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 16 2014

Application
Western blot
Loading amount
10000 cells
Gel Running Conditions
Reduced Denaturing (12%)
Sample
Mouse Cell lysate - whole cell (Mouse Glioblastoma (GL261))
Specification
Mouse Glioblastoma (GL261)
Treatment
HDAC inhibitors for 24hrs
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 28°C
Username

Abcam user community

Verified customer

Submitted Sep 12 2014

Application
Immunoprecipitation
Immuno-precipitation step
Other - Protein G Dynabeads
Sample
Human Cell lysate - other (H23)
Specification
H23
Total protein in input
250 µg
Username

Abcam user community

Verified customer

Submitted Dec 18 2013

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Sample
Human Cell (U87 human primary Glioblastoma)
Specification
U87 human primary Glioblastoma
Permeabilization
Yes - 0.3% triton X 100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 11 2013

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Neuro2A, mouse neuroblastoma)
Specification
Neuro2A, mouse neuroblastoma
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 11 2013

The FACS data on the datasheet was from cells stained with a purified version of the antibody for which the concentration was known to be 0.4 mg/ml. The antibody is currently produced as ascites fluid. The amount of specific antibody in ascites is typi...

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