Anti-alpha Tubulin antibody [TU-01] (ab7750)

Overview

  • Product nameAnti-alpha Tubulin antibody [TU-01]See all alpha Tubulin primary antibodies ...
  • Description
    Mouse monoclonal [TU-01] to alpha Tubulin
  • Tested applicationsFlow Cyt, IHC-P, ELISA, ICC, ICC/IF, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Cow, Dog, Human, Pig
    Predicted to work with: Rat, Chicken, Xenopus laevis, Plants, a wide range of other species, all Mammals, Xenopus tropicalis
  • Immunogen

    Full length native protein (purified) corresponding to Pig alpha Tubulin aa 37-154.

  • Epitopeaa 65-97 on N-terminal structural domain
  • Positive control
    • This antibody gave a positive signal in IF/ICC in HeLa cell line. Western Blot Positive control Pig Brain Lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab7750 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 1µg for 106 cells.
IHC-P Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ELISA Use at an assay dependent concentration.
ICC 1/100. For ICC, we recommend not only fixing but also permeabilizing the cells.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 55 kDa.

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 [TU-01] images

  • All lanes : Anti-alpha Tubulin antibody [TU-01] (ab7750) at 1/500 dilution

    Lane 1 : Dog 1 heart whole tissue lysate (left ventricle)
    Lane 2 : Dog 2 heart whole tissue lysate(left ventricle)
    Lane 3 : Dog 3 heart whole tissue lysate (left ventricle)
    Lane 4 : Dog 4 heart whole tissue lysate (left ventricle)

    Lysates/proteins at 80 µg per lane.

    Secondary
    HRP conjugated sheep polyclonal antibody
    developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 55 kDa (why is the actual band size different from the predicted?)


    Exposure time : 20 minutes

    This image is courtesy of an Abreview submitted by Dr sudhish mishra

    See Abreview

  • ICC/IF image of ab7750 stained HeLa cells. The cells were 4% formaldehyde 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 (ab7750, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) 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.
  • Ab7750 staining human normal skin. Staining is localised to the cytoplasm.
    Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system at room temperature. Sections were rehydrated and antigen retrieved with a retrieval buffer EDTA pH 9.0 . Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with an amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

  • Overlay histogram showing HEK293 cells stained with ab7750 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton 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 (ab7750, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Triton used under the same conditions.

References for Anti-alpha Tubulin antibody [TU-01] (ab7750)

This product has been referenced in:
  • Kozhemyakina E  et al. GATA6 is a crucial regulator of Shh in the limb bud. PLoS Genet 10:e1004072 (2014). WB . Read more (PubMed: 24415953) »
  • Pastorelli L  et al. Epithelial-derived IL-33 and its receptor ST2 are dysregulated in ulcerative colitis and in experimental Th1/Th2 driven enteritis. Proc Natl Acad Sci U S A 107:8017-22 (2010). WB ; Human . Read more (PubMed: 20385815) »

See all 8 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
Sample Plants Cell (meiocyte)
Specification meiocyte
Permeabilization Yes - Triton X-100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 23 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - other (U2OS)
Specification U2OS
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Mar 17 2014

Thank you for contacting us.

The following antibody should be working with plant samples;

http://www.abcam.com/alpha-tubulin-antibody-tu-01-ab7750.html

I hope this information is helpful to you. Please do not hesitate to cont...

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Thank you for contacting us and reporting the problems you have encountered in using the anti-alpha Tubulin antibody [TU-01] (ab7750).

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (HEK 293T)
Total protein in input 350 µg
Specification HEK 293T
Immuno-precipitation step Protein A/G
Username

Abcam user community

Verified customer

Submitted Jan 26 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Hek 293)
Loading amount 10 µg
Specification Hek 293
Gel Running Conditions Reduced Denaturing (10%)
Blocking step (agent) for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 22°C
Username

Dr. Chris Tracy

Verified customer

Submitted Jan 12 2012

Thank you for your enquiry and your interest in our products. Anti-alpha tubulin (clone TU-01) can be used as loading control together with antibody HLA Class I (ab23755) . Currently, we have several forms of this antibody ie. unconjugated as well as c...

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Thank you for your enquiry. I can confirm that the loading control protein alpha tubulin should only be located in the cytoplasm. We have several antibodies for detection of alpha tubulin which have been tested in WB, human and pig. I can recommend ...

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I am sorry for the delayed response. We have not tested it in Fungi but we would expect it to react with Fungi. The secondary goat anti-mouse is compatible with ab7750. I hope this helps. Please do not hesitate to contact us if you have any further ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (forebrain)
Loading amount 10 µg
Specification forebrain
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 19 2007

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