Product nameAnti-Tubulin antibody [YOL1/34] - Microtubule Marker
See all Tubulin primary antibodies
DescriptionRat monoclonal [YOL1/34] to Tubulin - Microtubule Marker
Tested applicationsSuitable for: WB, ELISA, IHC-FoFr, RIA, Flow Cyt, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Dog, Human, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Alligator
Predicted to work with: Plants, a wide range of other species, Mammals
Full length native protein (purified) corresponding to Saccharomyces cerevisiae Tubulin.
Epitopeab6161 binds to an epitope between amino acids 414 and 422 of alpha tubulin.
- WB: HeLa and NIH3T3 whole cell lysates and rat brain tissue lysate. Flow Cyt: methanol fixed/tween permeabilised HeLa cells. ICC/IF: HeLa, NIH/3T3 and human macrophage cells.
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.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
Primary antibody notesHas been used for the selection of specific recombinant antibodies engineered to incorporate its epitope. It is also useful for studying the function of microtubules.
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.
|WB||Use a concentration of 1 µg/ml.|
|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.|
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.
modificationsUndergoes 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
- Tubulin beta 2b antibody
- Alpha tubulin antibody
- Alpha-tubulin ubiquitous antibody
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: 20
µg/lane HeLa (Human) whole cell lysates (ab7898).
Lane 2: 20
µg/lane 3T3 (Mouse) whole cell lysate (ab7901).
Lane 3: 20
µg/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
All lanes : Yeast (Saccharomyces cerevisiae) whole cell extract prepared by bead-beating
Lysates/proteins at 5 µg per lane.
All lanes : 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
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
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.
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.
This product has been referenced in:
- Gao K et al. SUMO peptidase ULP-4 regulates mitochondrial UPR-mediated innate immunity and lifespan extension. Elife 8:N/A (2019). Read more (PubMed: 30642431) »
- Lo YH et al. Cryo-EM structure of the essential ribosome assembly AAA-ATPase Rix7. Nat Commun 10:513 (2019). Read more (PubMed: 30705282) »