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Technical Data Sheet


Actin & Tubulin
Antibody Sampler 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
AM2021 Dutta, P et al. (2014) J Neurochem. 130(3):360 WB: rat brain
AM2021 Muirhead, G et al. (2014) J Mol Neurosci. 53(1):125 WB: rat brain
AM2021 Pritchard, AJ et al. (2014) PLoS One. 9(6):e99444 WB: mouse splenocytes
TM1541 Wang, Q. et al. (2014) J Virol. 88(9):4853 WB: HeLa
AP1671 Vonach, C. et al. (2011) British J Cancer. 105:263. WB: human endothelial 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 actin and tubulin antibody sampler kit can be used to examine phosphorylation of actin (Tyr-53) and tubulin (Ser-172). In addition, the antibodies included in the kit can be used to detect actin and tubulin expression patterns by western blot or immunocytochemistry.

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).


Immunocytochemical labeling using anti-Actin (N-terminal) and anti-Actin (Tyr-53) polyclonal antibodies in C2C12 cells control (left) or treated with pervanadate (1 mM) for 30 min (middle). The cells were fixed in paraformaldehyde and permeabilized in acetone. Both antibodies were used in the presence of blocking peptide: Actin (N-terminal) peptide (AX1655) or phospho-Actin (Tyr-53) peptide (AX1675), respectively (right).

Microtubules (MTs) and actin filaments are cytoskeletal elements that play an essential role in cell division, cytoplasmic organization, cell morphology, and motility. 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. Actin filaments are polymers formed by one of six different actin isoforms found in vertebrates. Actin exists in two principal forms, globular, monomeric (G) actin, and filamentous polymeric (F) actin. Phosphorylation may regulate the activity tubulin and actin isoforms. Cdk1 phosphorylation of Ser-172 in β-tubulin occurs in mitotic cells and may impair tubulin incorporation into microtubules. In Dictyostelium, actin phosphorylation at Tyr-53 occurs in response to cell stress and this phosphorylation may alter actin polymerization. In B cells, SHP-1 tyrosine dephosphorylation of actin leads to actin filament depolymerization following BCR stimulation.

Background References
Diaz-Nido, J. et al. (1990) J Biol. Chem. 265(23):13949.
Jungbluth, A. et al. (1995) FEBS Let. 375:87.
Fanarraga, M.L. et al. (1999) Eur. J. Neurosci. 11:517.
Westermann, S. & Weber, K. (2003) Nat. Rev. Mol. Cell. Biol. 4:938.
Baba, T. et al. (2003) J. Immunol. 170: 3762.
Winder, S.J. et al. (2005) J. Cell Sci. 118:651.
Liu, X. et al. (2006) Proc Nat Acad Sci U S A. 103(37):13694.
Fourest-Lieuvin, A. et al. (2006) Mol. Biol. Cell. 17(3):1041.
Buffer and Storage
Mouse monoclonal and rabbit polyclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store all at –20°C. Stable for 1 year.
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