Overview

  • Product name
    Anti-alpha Tubulin antibody [TU-01]
    See all alpha Tubulin primary antibodies
  • Description
    Mouse monoclonal [TU-01] to alpha Tubulin
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow 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, Mammals, Xenopus tropicalis
  • Immunogen

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

  • Epitope
    aa 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
  • General notes

    This product was changed from ascites to tissue culture supernatant on 24th January 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

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.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

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

  • 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 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
    All lanes : 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

    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 anti-mouse DyLight® 488 (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.

  • 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 anti-mouse DyLight® 488 (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.

  • Immunocytochemistry/ Immunofluorescence analysis of NIH 3T3 (mouse embryonal fibroblast) cells labelling alpha tubulin (green) with ab7750. Vimentin was stained red as counterstain. DAPI was used to stain the nuclei blue. 

References

This product has been referenced in:
  • Refolo V  et al. Progressive striatonigral degeneration in a transgenic mouse model of multiple system atrophy: translational implications for interventional therapies. Acta Neuropathol Commun 6:2 (2018). Read more (PubMed: 29298733) »
  • Wei J  et al. Anti-proliferative effect of isorhamnetin on HeLa cells through inducing G2/M cell cycle arrest. Exp Ther Med 15:3917-3923 (2018). Read more (PubMed: 29563987) »
See all 20 Publications for this product

Customer reviews and Q&As

1-10 of 19 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (oral epithelial)
Specification
oral epithelial
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Dr. Tina Yuan

Verified customer

Submitted Sep 26 2018

Application
Western blot
Sample
Plants Tissue lysate - whole (Nicotiana benthamiana)
Gel Running Conditions
Non-reduced Denaturing (4-20% precut gels)
Loading amount
10 µg
Treatment
lysis buffer 12h
Specification
Nicotiana benthamiana
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 7.5% · Temperature: 20°C

Huan Zhang

Verified customer

Submitted Nov 13 2017

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

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

Abcam user community

Verified customer

Submitted Mar 17 2014

Question
Answer

Thank you for contacting us.

The following antibody should be working with plant samples;

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

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

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 very seriously, and investigate every product that we feel may not be performing correctly.

As tubulin is the major building block of microtubules and is present in most eukaryotic cells there is a wide range of samples you could use as a positive control. If you have any human or mouse cell lines available such as HeLa or NIH-3T3 these should work well. Otherwise most tissue samples should also be positive, we have used both heartand skin tissue with this particular antibody.

If you would like I can take a look at the protocol you have been using and the results you have found in order to see if there are any changes I could suggest in order to improve the results observed so far. If you would like for me to do this, please could you fill out thequestionnaire I have attached to this email. If you could include any images you have of the blots performed that would be very helpful.

I look forward to receiving your reply.

Read More
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

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

Dr. Chris Tracy

Verified customer

Submitted Jan 12 2012

Answer

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 conjugated: - ab7750 - Anti-alpha Tubulin antibody [TU-01] - ab18247 - Anti-alpha Tubulin antibody [TU-01] (FITC) - ab46708 - Anti-alpha Tubulin antibody [TU-01] (Dyomics® 547) - ab74696 - Anti-alpha Tubulin antibody [TU-01] (Biotin) - ab78108 - Anti-alpha Tubulin antibody [TU-01] I hope this helps and if I can assist further, please do not hesitate to contact me.

Read More

Answer

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 to review the datasheets for further information. ab15246 Anti-alpha Tubulin antibody Rabbit polyclonal Applications: ICC/IF, IHC-Fr, IHC-P, IP, WB Reacts with: Fruit fly (Drosophila melanogaster), Human, Mouse, Pig, Rat, Zebrafish Click here (or use the following: https://www.abcam.com/index.html?datasheet=15246). ab7750 Anti-alpha Tubulin antibody [TU-01] Mouse monoclonal Applications: ELISA, Flow Cyt, ICC, IF, IHC-P, IP, WB Reacts with: Human, Mouse, Arabidopsis thaliana, Cow, Dog, Pig Click here (or use the following: https://www.abcam.com/index.html?datasheet=7750). ab80779 Anti-alpha Tubulin antibody [DM1A] - BSA and Azide free Mouse monoclonal Applications: EM, Flow Cyt, ICC/IF, IHC-P, IP, WB Reacts with: Human, Mouse, Rat, Chicken, Cow, Gerbil, Guinea pig, Pig, Xenopus laevis Click here (or use the following: https://www.abcam.com/index.html?datasheet=80779). If you require any further assistance to find a suitable nuclear marker, please do not hesitate to let me know. I hope this will be helpful to you. Should you have any further questions, please do not hesitate to contact us.

Read More

1-10 of 19 Abreviews or Q&A

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