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

Mouse monoclonal alpha Tubulin (acetyl K40) antibody [6-11B-1]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Sheep, Human, Monkey, Sea urchin. Cited in 106 publication(s).

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)
  • Host species
    Mouse
  • 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
    All lanes : 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:
  • Choi YJ  et al. Mutations of ADAMTS9 Cause Nephronophthisis-Related Ciliopathy. Am J Hum Genet 104:45-54 (2019). Read more (PubMed: 30609407) »
  • Wormser O  et al. SCAPER localizes to primary cilia and its mutation affects cilia length, causing Bardet-Biedl syndrome. Eur J Hum Genet N/A:N/A (2019). Read more (PubMed: 30723319) »
See all 109 Publications for this product

Customer reviews and Q&As

1-10 of 15 Q&A

Answer

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 typically 1 - 10 mg/ml.

A volume of 1.0 ul per 10e6 cells is a standard starting point for determining the optimal dilution of ascites for FACS, but you may need to adjust the amount up or down within a range of, for instance, 0.25 - 4 ul, depending on your initial results.

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Answer


We would recommend using a pH6 sodium citrate retrieval.

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Question
Answer

Thank you for your enquiry.

I can confirm that ab24610 antibody is sold as ascites. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet. Antibody concentration is usually determined by protein assay, and serum / ascites / tissue culture supernatant will contain a lot of other proteins, which means the antibody quantification would not be accurate.

I can confirm that for ascites, concentration of antibody is known to very between 5 - 10 mg/ml.

I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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Answer

Thank you for your reply. I will be taking care of your query as Carolyn is out of the office.

I don't know enough about acetylated alpha Tubulin expression in primary cilium to be able to tell you for certain whether or not it is specific for primary cilium. Plus we do not have a protocol for ab24610 for specifically staining primary cilium. However, I did find that this clone has been cited as a marker for primary cilia in this article:

http://jcb.rupress.org/content/178/3/363.full

A quick literature search also suggests Adenyl Cyclase 3 (we carry ab125093 for this protein) may be a good marker for primary cilium. I'd recommend looking in the literature for more specific protocol recommendations.

http://www.biomedcentral.com/1471-2202/12/71

https://www.abcam.com/ADCY3-antibody-ab125093.html

I hope this information helps. Please contact us with any other questions.

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Answer

Thank you for your patience. After checking with the lab, we are not completely sure what the bright green staining is. We suspect that these are recently divided cells and the staining we are seeing is clumps of tubulin as the cells are pulling apart. I hope this helps, please let me know if you need any additional information or assistance.

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Answer

Thank you for contacting Abcam regarding ab24610.


Regarding the specificity of this antibody, it will recognize structures containing acetylated alpha-tubulin. In terms of structures stained, acetylated alpha tubulin is present in various microtubule structures and plays a role in stabilizing the structures of all microtubules.We expect the antibody will bind to primary cilia, centrioles, mitotic spindles, midbodies and to subsets of cytoplasmic microtubules in 3T3 and HeLa cells.



I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

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Answer

Thank you for contacting Abcam.

The Ascites storage buffer does not contain Sodium Azide and so you do not have to worry about that.

If there is anything else I can help you with, please let me know.

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Question
Answer

DISCOUNT CODE: *
Expiration date: *


I am very pleased to hear you would like to accept our offer and test ab24610 in Xenopus Laevis. This code will give you: 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for Xenopus Laevis and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting us and sorry for the delay in getting back to you.

I am now able to confirm that these antibodies are produced from the same clone 6-11B-1 and should produce very similar results. We have them separated into two different products as they are sourced from different places and this also means that we have slightly different information regarding their uses in different applications and species they have been used in.

I am sorry if the information has been confusing. If you have any further questions, please do not hesitate to contact us again.

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Question
Answer

Vielen Dank für Ihren Anruf.

Ich habe nun vom Labor die Antwort bekommen, dass der ab24610 leider noch nicht Verkaufsbereit ist. Unglücklicherweise, hatten wir viele Probleme mit diesem Antikörper und deshalb wird der Klon momentan gerade "subcloned". Falls ein Subklon dann positive ist, wird die Produktion aufgefahren. Auch falls alle Etappen positive ausfallen, wird dies jedoch mehrere Wochen dauern.

Es tut mir leid, Ihnen keine besseren Nachrichten geben zu können momentan. Ich kann Ihnen empfehlen, sich auf dem Datenblatt unter "email me" einzutragen. So werden Sie benachrichtigt, sobald der Antikörper wieder erhältlich ist. Click here (or use the following: https://www.abcam.com/index.html?datasheet=24610).

Bitte entschuldigen Sie all die Umstände. Bitte zögern Sie auch nicht, uns wieder zu kontaktieren, sollten Sie weitere Fragen oder Bedenken haben.

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1-10 of 15 Q&A

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