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Technical Data Sheet


Microtubule Labeling
Immunocytochemistry Kit


Kit Components
Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Product References
TP1691 Sitaras, N. et al. (2015) Am J Pathol. 185(2):581 IF: mouse retinal ganglion cells
TM1541 Wang, Q. et al. (2014) J Virol. 88(9):4853 WB: HeLa
TM4111 Gehrig, S.M. et al. (2012) Nature. 484(7394):394. WB: mouse skeletal muscle
TP1691 Saphieha, P. et al. (2012) Nutr Diabetes. 2:36. ICC: mouse retina cells
TM1541 Meacham, CE. et al. (2011) Nature Genetics. 41:1133. WB: mouse lymphoma
TM1541 Meshki, J. et al. (2011) PLoS ONE. 6(9):e25332. WB: U3&3MG
TM1541 Campbell, S. et al. (2010) J Biol Chem. 285:4695. WB: human HeLa cells
Kit Summary
The microtubule lableing kit can be used for immunocytochemical labeling of tubulin. The kit includes monoclonal antibodies to α- and βI-tubulin, and a rabbit polyclonal antibody to βIII-tubulin. The kit also includes goat-anti-rabbit conjugated to DyLight® 594 and goat anti-mouse conjugated to DyLight® 488 for dual labeling experiments.

Immunocytochemical labeling of Tubulin isoforms. In rat A7r5 cells, microtubules were labeled using anti-βI-Tubulin (TM1541) (left) and anti-α-tubulin (TM4111) (right). In NGF-differentiated rat PC12 cells, microtubules were labeled using anti-βIII-Tubulin (TP1691) (middle). The antibodies were detected using the appropriate secondary antibody conjugated to DyLight® 488 or 594.

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 α- and β-Tubulin isoforms. Multiple kinases can phosphorylate Ser-444 at the C-terminus of βIII-Tubulin in vitro, and unphosphorylated Ser-444 may be an early marker for cells of neuronal lineage. Cdk1 can phosphorylate Ser-172 in β-Tubulin during mitosis and this may impair tubulin incorporation into microtubules. In α-tubulin, PKC can phosphorylate multiple residues, and phosphorylation of Ser-165 by PKC has been implicated in promoting cell motility.

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.
Abeyweera, T.P. et al. (2009) J Biol Chem. 284(26):17648.
Buffer and Storage
Mouse monoclonal, rabbit polyclonal, and secondary reagents are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
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