Anti-alpha Tubulin antibody [4G1] (ab28439)
- Product nameAnti-alpha Tubulin antibody [4G1]See all alpha Tubulin primary antibodies ...
- DescriptionMouse monoclonal [4G1] to alpha Tubulin
- Tested applicationsELISA, WB, Flow Cyt, ICC/IF more details
- Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide corresponding to C terminal amino acids of Human alpha Tubulin, conjugated to KLH
- Positive control
- SH-SY5Y cell lysate This antibody gave a positive result when used in the following methanol fixed cell lines: HeLa.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: None
Constituents: 50% Glycerol, PBS
- Concentration information loading...
- PurityIgG fraction
- Clonality Monoclonal
- Clone number4G1
- Light chain typekappa
- Research Areas
Our Abpromise guarantee covers the use of ab28439 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||ELISA: Use at an assay dependent concentration.|
|WB||WB: 1/2000 - 1/10000. Predicted molecular weight: 54 kDa.|
|Flow Cyt||Flow Cyt: Use 1µg for 106 cells.|
|ICC/IF||ICC/IF: Use a concentration of 10 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.
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.
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Anti-alpha Tubulin antibody [4G1] images
ab28439 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 ab28439 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) 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. This antibody also gave a positive result in Methanol fixed (100%) Hek293 and MCF-7 cells, and also in formaldehyde fixed (4%) HeLa, Hek293, HepG2, and MCF-7 cells at 10ug/ml.
All lanes : Anti-alpha Tubulin antibody [4G1] (ab28439) at 1/2000 dilution
Lane 1 : SH-SY5Y cell lysate
Lane 2 : Mouse brain tissue
Lane 3 : Rat brain tissue
Predicted band size : 54 kDa
Observed band size : 54 kDa
Overlay histogram showing HeLa cells stained with ab28439 (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 (ab28439, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
References for Anti-alpha Tubulin antibody [4G1] (ab28439)
This product has been referenced in:
- Hao J et al. Dorsomorphin, a selective small molecule inhibitor of BMP signaling, promotes cardiomyogenesis in embryonic stem cells. PLoS One 3:e2904 (2008). WB ; Mouse . Read more (PubMed: 18682835) »