Anti-Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161)

Overview

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab6161 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
WB Use a concentration of 1 µg/ml.
ELISA 1/4000.
IHC-FoFr 1/600. PubMed: 15831501Suggested working dilution of 1/600.
RIA Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.
ICC/IF Use a concentration of 5 µg/ml.

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
    Undergoes a tyrosination/detyrosination cycle, the cyclic removal and re-addition of a C-terminal tyrosine residue by the enzymes tubulin tyrosine carboxypeptidase (TTCP) and tubulin tyrosine ligase (TTL), respectively.
    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-tubulins 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
    • Tubulin beta 2b antibody
    • Alpha tubulin antibody
    • Alpha-tubulin ubiquitous antibody
    • beta Ib tubulin antibody
    • CDCBM5 antibody
    • CDCBM6 antibody
    • fd02b12 antibody
    • K ALPHA 1 antibody
    • M40 antibody
    • OK/SW-cl.56 antibody
    • TBA1B_HUMAN antibody
    • TUBA1B antibody
    • TUBB1 antibody
    • TUBB2 antibody
    • TUBB5 antibody
    • tubulin alpha 1b antibody
    • Tubulin alpha-1B chain antibody
    • Tubulin alpha-ubiquitous chain antibody
    • Tubulin beta 1b antibody
    • Tubulin beta 2A antibody
    • tubulin beta 2A class IIa antibody
    • Tubulin beta antibody
    • tubulin beta chain antibody
    • tubulin beta class I antibody
    • tubulin beta-1 chain antibody
    • tubulin beta-2A chain antibody
    • tubulin beta-5 chain antibody
    • Tubulin K-alpha-1 antibody
    • tubulin, alpha, ubiquitous antibody
    • tubulin, beta 2A class IIa antibody
    • tubulin, beta polypeptide 2 antibody
    • tubulin, beta polypeptide antibody
    • tubulin, beta, class IIA antibody
    • wu:fd02b12 antibody
    • zgc:55461 antibody
    see all

Anti-Tubulin antibody [YOL1/34] - Microtubule Marker images

  • Western blot against tubulin with ab6161 at 1/3000.  Secondary Rabbit anti-Rat IgG HRP (ab6734)was used at 1/2000.  Exposure time: 2mins.

    Lane 1: 20µg/lane HeLa (Human) whole cell lysates (ab7898).

    Lane 2: 20µg/lane 3T3 (Mouse) whole cell lysate (ab7901).

    Lane 3: 20µg/lane Rat brain tissue lysate (ab7942).

  • Confocal image of  21 day in vitro rat hippocampal neurons, stained with rat monoclonal antibody to Tubulin - Microtubule Marker (ab6161) in green at 1/500 and Microtubule Associated protein 2 in blue.

    This picture was kindly supplied as part of the review submitted by Dr Jonathon Burman.

  • Overlay histogram showing HeLa cells stained with ab6161 (red line). The cells were fixed with 80% methanol (5 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 (ab6161, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (ab18450, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • ICC/IF image of ab6161 stained Hela cells. The cells were 100% methanol fixed (5 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 (ab6161, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rat- H&L, pre-adsorbed (ab98420) 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.

  • Cultured human macrophages were used with ab6161 at 1/1000 for immunofluorescence. Cells were fixed with cold 2% formaldehyde for 20mins.

    Green staining is Alexa 568, Blue staining is DAPI stain.

    This cell represents a young macrophage, the staining patterns varied as the cells aged in culture.

  • All lanes : Anti-Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161) at 1/2000 dilution

    Lane 1 : Yeast whole cell extract prepared by bead-beating
    Lane 2 : Yeast whole cell extract prepared by bead-beating
    Lane 3 : Yeast whole cell extract prepared by bead-beating
    Lane 4 : Yeast whole cell extract prepared by bead-beating
    Lane 5 : Yeast whole cell extract prepared by bead-beating
    Lane 6 : Yeast whole cell extract prepared by bead-beating
    Lane 7 : Yeast whole cell extract prepared by bead-beating
    Lane 8 : Yeast whole cell extract prepared by bead-beating
    Lane 9 : Yeast whole cell extract prepared by bead-beating
    Lane 10 : Yeast whole cell extract prepared by bead-beating

    Lysates/proteins at 5 µg per lane.

    Secondary
    HRP conjugated goat anti-rat antibody
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 50 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview

  • ICC/IF image of ab6161 stained human HepG2 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab6161, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rat IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, HEK 293 and MCF7 cells.

  • Anti-Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161) at 1 µg/ml + Brain (Rat) Tissue Lysate at 10 µg

    Secondary
    Rabbit polyclonal to Rat IgG - H&L (HRP) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 54 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 minutes
  • ab6161 staining mouse NIH 3T3 fibroblast cells by ICC/IF. Cells were PFA fixed  and permeabilized in 0.2% Triton X-100 prior to blocking in 5% BSA for 45 minutes at RT.  The primary antibody was diluted 1/1000 and incubated with the sample for 1 hour.  An Alexa Fluor® 568 conjugated goat anti-rat antibody, diluted 1/3000, was used as the secondary.

     

    See Abreview

  • ab6161 staining tubulin HeLa cells treated with anisomycin (ab120495), by ICC/IF. Increase in tubulin expression correlates with increased concentration of anisomycin as described in literature.
    The cells were incubated at 37°C for 6h in media containing different concentrations of ab120495 (anisomycin) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab6161 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rat polyclonal antibody (ab98386) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

References for Anti-Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161)

This product has been referenced in:
  • Keller P  et al. An Antifungal Benzimidazole Derivative Inhibits Ergosterol Biosynthesis and Reveals Novel Sterols. Antimicrob Agents Chemother 59:6296-307 (2015). ICC/IF ; Candida albicans . Read more (PubMed: 26248360) »
  • Vaithiyalingam S  et al. Insights into Eukaryotic Primer Synthesis from Structures of the p48 Subunit of Human DNA Primase. J Mol Biol 426:558-69 (2014). WB ; Saccharomyces cerevisiae . Read more (PubMed: 24239947) »

See all 43 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Zebrafish Cell (Embryo)
Permeabilization Yes - Methanol
Specification Embryo
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative Paraformaldehyde
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Submitted Oct 20 2016

Application Western blot
Sample Saccharomyces cerevisiae Cell lysate - whole cell (BY4741)
Gel Running Conditions Reduced Denaturing (4%-15%)
Loading amount 10 µg
Specification BY4741
Blocking step Milk as blocking agent for 15 minute(s) · Concentration: 2% · Temperature: 25°C
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Submitted Dec 10 2015

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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Application Western blot
Sample Leishmania Cell lysate - whole cell (Leishmania)
Loading amount 10 µg
Specification Leishmania
Gel Running Conditions Reduced Denaturing (gel 10%)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Nov 17 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - other (Non-small cell lung carcinoma)
Loading amount 10 µg
Specification Non-small cell lung carcinoma
Gel Running Conditions Reduced Denaturing (4-12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Dec 01 2010

Application Western blot
Sample Candida albicans Cell lysate - whole cell (Candida Albicans Whole Cell Prep)
Loading amount 5e+007 cells
Specification Candida Albicans Whole Cell Prep
Gel Running Conditions Non-reduced Denaturing (10% SDS PAGE)
Blocking step LI-COR Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 25°C
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Submitted Oct 07 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (huh-7 cells)
Loading amount 10 µg
Specification huh-7 cells
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Jul 01 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (primary macrophages)
Specification primary macrophages
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Apr 16 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Methanol
Permeabilization No
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 27°C
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Submitted Dec 01 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Methanol
Permeabilization Yes - 0.1% Triton X-100
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
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Submitted Nov 14 2008

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