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Technical Data Sheet

TP1811

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unphosphorylated βIII-Tubulin (Ser-444)
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
50 kDa

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Western blot analysis of mouse brain. The blot was probed with anti-unphosphorylated βIII-Tubulin (Ser-444) (lanes 1-3) and anti-βIII-Tubulin (C-terminus) (lanes 4-6) polyclonal antibodies. Both antibodies were used in the presence of unphosphorylated βIII-Tublin (Ser-444) peptide (lanes 2 & 5; TX1815) and phospho-βIII-Tublin (Ser-444) peptide (lanes 3 & 6; TX1695).

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 a/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
Unphosphorylated βIII-Tubulin (Ser-444) synthetic peptide (coupled to KLH) corresponding to amino acid residues around serine 444 of human βIII-Tubulin. This sequence is not found in bI or βII-Tubulin isotypes, but is well conserved in βIII-Tubulins from rat and mouse.
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 phospho-βIII-Tubulin (Ser-444) peptide before affinity purification using unphosphorylated βIII-Tubulin (Ser-444) peptide (without carrier). The antibody detects a 50 kDa* protein corresponding to the molecular mass of unphosphorylated βIII-Tubulin on SDS-PAGE immunoblots of purified brain tubulin and mouse brain tissue.

*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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TP1781
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