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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, WB more details
  • Species reactivity
    Reacts with: Mouse, Cow, Dog, Human, Pig, Arabidopsis thaliana
    Predicted to work with: a wide range of other species, all Mammals
  • Immunogen

    Fraction of tubulin purified from pig brain by two cycles of polymerization-depolymerization.

  • Epitopeaa 65-97 on N-terminal structural domain
  • Positive controlThis antibody gave a positive signal in IF/ICC in HeLa cell line.

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

Target

  • Alternative names
      Alpha tubulin 2 antibodyAlpha tubulin 3 antibodyAlpha tubulin 3C/D antibody
      Alpha tubulin ubiquitous antibodyB ALPHA 1 antibodybA408E5.3 antibodyH2 ALPHA antibodyHum a tub1 antibodyHum a tub2 antibodyLIS3 antibodyTUBA1 antibodyTUBA1A antibodyTUBA2 antibodyTUBA3 antibodyTUBA3C antibodyTUBA3D antibodyTUBA4A antibodyTubulin alpha 1 chain antibodyTubulin alpha 1A chain antibodyTubulin alpha 2 chain antibodyTubulin alpha 3 chain antibodyTubulin alpha 3C/D chain antibodyTubulin alpha antibodyTubulin alpha ubiquitous chain antibodyTubulin B alpha 1 antibodyTubulin K alpha 1 antibodyTubulin, alpha 1 (testis specific) antibodyTubulin, alpha 1a antibodyTubulin, alpha 2 antibodyTubulin, alpha 3 antibodyTubulin, alpha 3c antibodyTubulin, alpha 4a antibodyTubulin, alpha, brain specific 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

  • 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 DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. 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.

  • 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.

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

This product has been referenced in:
  • 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) »
  • Hemsley PA  et al. Assaying protein palmitoylation in plants. Plant Methods 4:2 (2008). WB . Read more (PubMed: 18190696) »

See all 7 Publications for this product

Product Wall

Displaying 1 - 10 of 15 results for Abreviews and Q&A

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

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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).

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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

Verified customer

Submitted Jan 26 2012

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

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 conj...

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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 to ...

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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 que...

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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

Verified customer

Submitted Jun 19 2007

Application Western blot
Sample Dog Tissue lysate - whole (Heart)
Loading amount 80 µg
Specification Heart
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Verified customer

Submitted Apr 27 2007

I have received more information about ab7750: The antibody was raised against pig brain microtubeule preparation. The epitope was mapped within 37-154 aa sequence (N-terminal domain). It reacts with wide variety of species including eukaryotes, howeve...

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