Overview

  • Product name
    Anti-Tubulin antibody [YL1/2]
    See all Tubulin primary antibodies
  • Description
    Rat monoclonal [YL1/2] to Tubulin
  • Host species
    Rat
  • Tested applications
    Suitable for: ELISA, IHC-Fr, IP, RIA, WB, Flow Cyt, ICC/IF, IHC (PFA fixed), IHC-P, IHC - Wholemount, IHC (Methanol fixed)more details
  • Species reactivity
    Reacts with: Mouse, Human, Pig, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, African green monkey
    Predicted to work with: a wide range of other species, Mammals
  • Immunogen

    Full length native protein (purified) corresponding to Saccharomyces cerevisiae Tubulin.

  • Epitope
    A linear sequence requiring an aromatic residue at the C terminus, with the two adjacent amino acids being negatively charged (represented by Glu-Glu-Tyr in Tyr-Tubulin).
  • Positive control
    • ICC/IF: HeLa cells. IHC-P: Human colon tissue. WB: HeLa, NIH/3T3, BALB/3T3 and PC-12 whole cell lysate. Flow Cyt: HeLa cells.
  • General notes

    This antibody clone is manufactured by Abcam.

    This antibody can be used as a loading control on Western blots (Allen et al.) and is not detected by anti-mouse Ig secondaries. It has been used in epitope tagging procedures to detect proteins tagged with a C-terminal Gly-Gly-Phe(OH) epitope. Under some circumstances this antibody may cross-react with other protein including E. coli rec A and oxidized actin.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab6160 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
ELISA Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
RIA Use at an assay dependent concentration.
WB 1/5000 - 1/10000.
Flow Cyt Use 1µg for 106 cells.

ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

ICC/IF 1/1000. (see PMID: 16230461)
IHC (PFA fixed) Use a concentration of 5 µg/ml. (from PubMed:16966421)
IHC-P Use at an assay dependent concentration.
IHC - Wholemount Use at an assay dependent concentration. PubMed: 25368174
IHC (Methanol fixed) 1/100. (from PubMed:16943269).

Target

  • Relevance
    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. Dimer of alpha and beta chains. A typical microtubule is a hollow water-filled tube with an outer diameter of 25 nm and an inner diameter of 15 nM. Alpha-beta heterodimers associate head-to-tail to form protofilaments running lengthwise along the microtubule wall with the beta-tubulin subunit facing the microtubule plus end conferring a structural polarity. Microtubules usually have 13 protofilaments but different protofilament numbers can be found in some organisms and specialized cells.
  • Cellular localization
    Cytoplasmic
  • Database links
    • Alternative names
      • Tubulin alpha-1 chain antibody
      • Tubulin alpha-3 chain antibody

    Images

    • Cytoskeleton and major extracellular matrix proteins in human DPSC (Dental pulp stem cell) were analyzed by immunofluorescence.

      Vimentin and tubulin (Panel D, control, E, (BD) and F, (BR)) are shown.

      After 7 days in contact with/without the materials (Biodentine (BD) and Bioroot (BR)), coated coverslip cultures were fixed in PBS (pH 7.4) containing 4% paraformaldehyde/5% sucrose for 10 minutes. For detection of intracellular molecules, the cells on the coverslips were permeabilized using 0.5% Triton X-100. To block background staining, cells were treated with PBS containing 1% BSA/1% glycine at 37°C for 20 minutes. Samples were incubated with the primary antibody at 4°C overnight or at 37°C for 2 hours. For double immunostaining, primary antibodies were incubated as above. Samples were then incubated with the appropriate secondary antibodies at 37°C for 1 hour. Cell nuclei were stained using DAPI.

      Scale Bar: 100 μm.

    • Egr3 localization is associated with microtubule organization in mouse oocytes.

      Mouse oocytes stained for Tubulin using ab6160 (Right panels, red) in ICC/IF.

      The localization of Egr3 and microtubule at thawing of vitrified oocytes. Vitrified MII oocytes were stored in LN2 for 2 weeks. Oocytes were taken out from LN2, incubated in decreasing concentrations of sucrose, and then fixed immediately. These oocytes were subjected to immunofluorescence staining with anti-Egr3 and ab6160 antibodies. Arrows indicate the growing arrays of microtubules at the site of Egr3 accumulation.

      Green: Egr3.

      Red: Microtubule (MT).

    • ICC/IF image of ab6160 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

      The cells were fixed in 100% methanol 5 minutes, permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6160, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was ab150165 Alexa Fluor® 488 goat anti-rat IgG (H+L) pre-adsorbed, used at a 1/1000 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

      The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.

      This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 minutes).

       

    • IHC image of Tubulin staining in human colon formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F.

      The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer pH 6 for 20 minutes. The section was then incubated with ab6160, 5 µg/ml, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • All lanes : Anti-Tubulin antibody [YL1/2] (ab6160) at 1 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 : PC-12 (Rat adrenal pheochromocytoma cell line) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Peroxidase Conjugated Rabbit Anti-Rat IgG (H+L) at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size: 50 kDa
      Observed band size: 52 kDa (why is the actual band size different from the predicted?)
      Additional bands at: 85 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 8 minutes
    • Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with ab6160 (red line).

      The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 (ab6160, 1 µg/1x106 cells) for 30 minutes at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (ab18450, 1 µg/1x106 cells) used under the same conditions.

      Acquisition of >5,000 events was performed.

    • Lanes 1 & 3 : Anti-Tubulin antibody [YL1/2] (ab6160) at 1/5000 dilution
      Lanes 2 & 4 : Anti-Tubulin antibody [YL1/2] (ab6160) at 1/10000 dilution

      Lanes 1-2 : HeLa (Human epithelial carcinoma cell line) whole cell lysate
      Lanes 3-4 : BALB/3T3 whole cell lysate (ab7901)

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Rabbit Anti-Rat IgG H&L (HRP) (ab6734) at 1/2000 dilution

      Predicted band size: 50 kDa
      Observed band size: 52 kDa (why is the actual band size different from the predicted?)
      Additional bands at: 17 kDa, 34 kDa, 80 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 10 seconds
    • ab6160 staining Tubulin in mouse trophoblast giant cells by ICC/IF (Immunocytochemistry/Immunofluorescence).

      Cells were fixed with methanol and blocked with 0.5% BSA for 30 minutes at 20°C. Samples were incubated with primary antibody (0.5% BSA, 0.1% Tween-20 PBS) for 1 hour at 20°C. An Alexa Fluor® 568 polyclonal Goat anti-Rat IgG (H+L) Cross-Adsorbed (1/750 dilution) was used as the secondary antibody.

      See Abreview

    • ab6160 staining Tubulin in mouse MEF cells by Immunocytochemistry/ Immunofluorescence.

      Cells were fixed with 2% PFA  and 96% Ethanol. Samples were incubated with primary antibody (1/2000 in 0.1% Saponin/1% BSA/PBS) for 1 hour. A Cyt3®-conjugated goat polyclonal to rat IgG (H&L) was used at dilution at 1/500 as secondary antibody.

      Red staining in the image represents Tubulin, whereas the green one resembles gamma-tubulin.

      See Abreview

    • ab6160 staining mouse prostate tissue sections by IHC-P.

      The tissue was serial sectioned at 6 microns, formaldehyde fixed and subjected to heat mediated antigen retrieval prior to blocking in 3% peroxidase for 5 minutes at 27°C. The primary antibody was diluted 1/500 and incubated with the sample for 16 hours at 4°C. A Cy5® conjugated goat anti-rat antibody was used as the secondary.

      See Abreview

    • ab6160 at 1/1000 dilution staining Tubulin in human WBC cells by Immunocytochemistry/ Immunofluorescence.

      Cells were fixed in acetone and then blocked in 5% serum for 1 hour at 25°C. No permeabilization was done. The primary antibody was used at 1/1000 dilution in PBS-Tween and incubated with sample at 4°C for 16 hours. An Alexa Fluor® 594 conjugated goat polyclonal to rat IgG was used as secondary at 1/500 dilution.

      See Abreview

    • This image was kindly supplied as part of the review submitted by Marko Kallio. ab6160 was used for immunofluorescence on male rat testis samples in order to visualize microtubules of meiotically deviding cells. The samples were fixed with 2% paraformaldehyde and 0.8% glutaraldehyde and the antibody was used at a dilution 1:2500 (red - tubulin, blue - DNA stained with DAPI).

    • ab6160 at a 1/200 staining Tubulin in mouse liver tissue sections by Immunohistochemistry (frozen sections) incubated for 9 hours at +4°C. Fixed in formaldehyde, permeabilized using 0.2% Triton X-100. Blocked using 2% BSA for 30 minutes at 20°C. Secondary used at a 1/200 dilution polyclonal Goat anti-rat IgG conjugated to Alexa Fluor® 555.

      See Abreview

    References

    This product has been referenced in:
    • Babcock HP Multiplane and Spectrally-Resolved Single Molecule Localization Microscopy with Industrial Grade CMOS cameras. Sci Rep 8:1726 (2018). Read more (PubMed: 29379074) »
    • Loison-Robert LS  et al. In vitro effects of two silicate-based materials, Biodentine and BioRoot RCS, on dental pulp stem cells in models of reactionary and reparative dentinogenesis. PLoS One 13:e0190014 (2018). Read more (PubMed: 29370163) »

    See all 200 Publications for this product

    Customer reviews and Q&As

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (293T cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Gradient Gel)
    Loading amount
    50 µg
    Specification
    293T cells
    Blocking step
    Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Sep 10 2018

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

    Abcam user community

    Verified customer

    Submitted Sep 06 2018

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (MEFs)
    Permeabilization
    Yes - Digitonin
    Specification
    MEFs
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 27°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Aug 31 2018

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (MEF cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Gradient Gel)
    Loading amount
    50 µg
    Specification
    MEF cells
    Blocking step
    Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Aug 01 2018

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (brain cortex)
    Gel Running Conditions
    Reduced Denaturing (8%)
    Loading amount
    30 µg
    Specification
    brain cortex
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 22°C
    Username

    Dr. Alexandre Magno

    Verified customer

    Submitted Jul 23 2018

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human cerebral organoid)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (12.5% gel)
    Loading amount
    20 µg
    Specification
    Human cerebral organoid
    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 May 30 2018

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Trophoblast giant cells)
    Permeabilization
    No
    Specification
    Trophoblast giant cells
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: 20°C
    Fixative
    Methanol
    Username

    Miss. Stephanie Chrysanthou

    Verified customer

    Submitted Jul 10 2017

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Tissue lysate - other (Hippocampus)
    Gel Running Conditions
    Reduced Denaturing (13)
    Loading amount
    6 µg
    Specification
    Hippocampus
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Dr. Sergi Bayod

    Verified customer

    Submitted Nov 28 2016

    Application
    IHC - Wholemount
    Sample
    Zebrafish Embryo (Early Embryo)
    Specification
    Early Embryo
    Username

    Abcam user community

    Verified customer

    Submitted Oct 27 2016

    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Sciatic nerve)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    20 µg
    Specification
    Sciatic nerve
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Mar 11 2016

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