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Technical Data Sheet

TP1721

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β-Tubulin (Ser-172), phospho-specific
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms, Ck
50 kDa

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Western blot analysis of purified brain tubulin untreated (lanes 1,3,5) or treated with ERK2 kinase to phosphorylate Ser-172 (lanes 2,4,6). The blot was probed with anti-β-Tubulin (a.a. 168-177) (lanes 1 & 2), anti-β-Tubulin (Ser-172) (lanes 3 & 4), and anti-β-Tubulin (TM1541) (lanes 5 & 6).

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Immunocytochemical labeling in C2C12 cells using anti-β-Tubulin (TM1541) monoclonal antibody and anti-β-Tubulin (Ser-172) polyclonal antibody. The specificity of the binding for the latter antibody was demonstrated by using the antibody in the presence of phospho-β-Tubulin (Ser-172) peptide (TX1725).

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Application
Dilution


ELISA
1:2000


ICC
1:100


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
Microtubules (MTs) are cytoskeletal elements that play an essential role in cell division and cytoplasmic organization. MTs are dynamic polymers of α/β-tubulin heterodimers. At least two populations of MTs, called dynamic and stable according to their rates of turnover, are readily distinguishable in cells. The proteins associated with MTs (MAPs) are among the best-known factors that regulate MT dynamics and stability. In addition, a variety of different post-translational modifications may also regulate MT dynamics and stability. Phosphorylation is one of these modifications and it can occur on serine, threonine, and tyrosine residues in β-Tubulin isoforms. Multiple kinases can phosphorylate Ser-444 at the C-terminus of βIII-Tubulin in vitro. Unphosphorylated Ser-444 in βIII-Tubulin is an early marker for cells of neuronal lineage, while phosphorylation of Ser-444 is upregulated after neuronal maturation and may preferentially occur in assembled MTs. By contrast, Cdk1 phosphorylation of Ser-172 in β-Tubulin occurs in mitotic cells and may impair tubulin incorporation into microtubules.


Background References
Diaz-Nido, J. et al. (1990) J Biol. Chem. 265(23):13949.
Fanarraga, M.L. et al. (1999) Eur. J. Neurosci. 11:517.
Westermann, S. & Weber, K. (2003) Nat. Rev. Mol. Cell. Biol. 4:938.
Fourest-Lieuvin, A. et al. (2006) Mol. Biol. Cell. 17(3):1041.
Immunogen
Phospho-βIII-Tubulin (Ser-172) synthetic peptide (coupled to KLH) corresponding to amino acid residues around serine 172 of human βIII-Tubulin. This sequence is identical to similar regions in βI, βII, and βIII-Tubulin isotypes, and is well conserved in tubulins from most eukaryotic species.
Buffer and Storage
Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
This antibody was cross-adsorbed to unphosphorylated βIII-Tubulin (Ser-172) peptide before affinity purification using phospho-βIII-Tubulin (Ser-172) peptide (without carrier). The antibody detects a 50 kDa* protein corresponding to the molecular mass of β-Tubulin on SDS-PAGE immunoblots of purified brain tubulin phosphorylated with ERK2 kinase.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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