Alpha6-Tubulin (Ser-165), phospho-specific polyclonal, anti-human, mouse, rat, chicken, fish

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 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 Ser-165 leading to increased cell motility in human breast cells.

Immunogen: Phospho-α6-Tubulin (Ser-165) synthetic peptide (coupled to KLH) corresponds to amino acid residues around serine 165 of human α6-Tubulin. This sequence is well conserved in most α-tubulin isoforms from most eukaryotic species, but is not conserved in β-Tubulin.

Specificity: This antibody was cross-adsorbed to unphosphorylated α6-Tubulin (a.a. 160-169) peptide before affinity purification using α6-Tubulin (Ser-165) peptide (without carrier). The antibody detects a 50 kDa* protein corresponding to the molecular mass of α-Tubulin on SDS-PAGE immunoblots of recombinant a1-tubulin phosphorylated with PKCα. This reactivity is not observed after lambda phosphatase treatment.

Application dilution:
ELISA: 1:2000
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 1 hour at room temperature.

Buffer/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.