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

Overview

  • Product name
    Anti-Tubulin antibody [YOL1/34] - Microtubule Marker
    See all Tubulin primary antibodies
  • Description
    Rat monoclonal [YOL1/34] to Tubulin - Microtubule Marker
  • Tested applications
    Suitable for: WB, ELISA, IHC-FoFr, RIA, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Alligator
    Predicted to work with: Plants, a wide range of other species, Mammal
  • Immunogen

    Full length native protein (purified) (S. cerevisiae).

  • Epitope
    ab6161 binds to an epitope between amino acids 414 and 422 of alpha tubulin.
  • Positive control
    • WB: HeLa and NIH3T3 whole cell lysates and rat brain tissue lysate. Flow Cyt: methanol fixed/tween permeabilised HeLa cells.
  • General notes

    This antibody clone is manufactured by Abcam.

    We can conjugate this antibody to FITC for you (please see ab150252 for details). This antibody can be used as a Western blotting loading control (Kops et al.) and as a Microtubule Marker.

    Has been used for the selection of specific recombinant antibodies engineered to incorporate its epitope. It is also useful for studying the function of microtubules.

     

Properties

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.

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

ICC/IF Use a concentration of 5 µg/ml.

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
    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 localization
    Cytoplasm > 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:
  • Beckmann BM RNA interactome capture in yeast. Methods 118-119:82-92 (2017). WB ; Saccharomyces cerevisiae . Read more (PubMed: 27993706) »
  • Guo W  et al. Decreased Human Leukocyte Antigen-G Expression by miR-133a Contributes to Impairment of Proinvasion and Proangiogenesis Functions of Decidual NK Cells. Front Immunol 8:741 (2017). WB . Read more (PubMed: 28702027) »

See all 65 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (NIH-3T3)
Permeabilization
Yes - 0.2% Triton X-100
Specification
NIH-3T3
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde
Username

Mr. Spencer Goodman

Verified customer

Submitted May 23 2017

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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

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...

Read More
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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

Submitted Dec 01 2008

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