Product nameAnti-alpha Tubulin antibody [TU-01]
See all alpha Tubulin primary antibodies
DescriptionMouse monoclonal [TU-01] to alpha Tubulin
Tested applicationsSuitable for: Flow Cyt, IHC-P, ELISA, ICC, ICC/IF, IP, WBmore details
Species reactivityReacts with: Mouse, Cow, Dog, Human, Pig
Predicted to work with: Rat, Chicken, Xenopus laevis, Plants, a wide range of other species, Mammals, Xenopus tropicalis
Full length native protein (purified) corresponding to Pig alpha Tubulin aa 37-154.
Epitopeaa 65-97 on N-terminal structural domain
- This antibody gave a positive signal in IF/ICC in HeLa cell line. Western Blot Positive control Pig Brain Lysate
This product was changed from ascites to tissue culture supernatant on 24th January 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
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: 0.097% Sodium azide
Concentration information loading...
Purification notesPurified from TCS. Purified by precipitation and chromatography. Purity >95% by SDS-PAGE.
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.
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-P||Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ELISA||Use at an assay dependent concentration.|
|ICC||1/100. For ICC, we recommend not only fixing but also permeabilizing the cells.|
|ICC/IF||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|WB||Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 55 kDa.|
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.
modificationsSome 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 chains 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
- Alpha-tubulin 1 antibody
- ALS22 antibody
- B ALPHA 1 antibody
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.
All lanes : 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
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 anti-mouse DyLight® 488 (ab96879) 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.
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 at room temperature. Sections were rehydrated and antigen retrieved with a retrieval buffer EDTA pH 9.0 . Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with an amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin. 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 anti-mouse DyLight® 488 (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.
Immunocytochemistry/ Immunofluorescence analysis of NIH 3T3 (mouse embryonal fibroblast) cells labelling alpha tubulin (green) with ab7750. Vimentin was stained red as counterstain. DAPI was used to stain the nuclei blue.
This product has been referenced in:
- Sudevan S et al. Mitochondrial dysfunction causes Ca2+ overload and ECM degradation-mediated muscle damage in C. elegans. FASEB J 33:9540-9550 (2019). Read more (PubMed: 31162948) »
- Wei J et al. Anti-proliferative effect of isorhamnetin on HeLa cells through inducing G2/M cell cycle arrest. Exp Ther Med 15:3917-3923 (2018). Read more (PubMed: 29563987) »