Catalog # AP1671

Actin (Tyr-53), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
ICC	1:50
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Ms, Ck

MW 42 kDa

Actin is a major cytoskeletal protein involved in diverse cellular functions including cell motility, adhesion, and morphology. Six different actin isoforms have been identified in vertebrates. There are four α isoforms: skeletal, cardiac, and two smooth muscle (enteric and aortic) actins, along with two cytoplasmic actins (β and γ). Actin exists in two principal forms, globular, monomeric (G) actin, and filamentous polymeric (F) actin. The assembly and disassembly of actin filaments, and also their organization into functional networks, is regulated by a variety of actin-binding proteins (ABPs). Phosphorylation may also be important for regulating actin assembly and interaction with ABPs. In Dictyostelium, phosphorylation of 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.

References

Liu, X. et al. (2006) Proc Nat Acad Sci U S A. 103(37):13694.
Winder, S.J. et al. (2005) J. Cell Sci. 118:651.
Baba, T. et al. (2003) J. Immunol. 170: 3762.
Jungbluth, A. et al. (1995) FEBS Let. 375:87.

Western blot analysis of mouse C2C12 cells untreated (lanes 1 & 3), or treated with pervanadate (1 mM) for 30 min (lanes 2 & 4). The blot was probed with anti-Actin (N-terminal) antibody (lanes 1 & 2) or anti-Actin (Tyr-53) antibody (lanes 3 & 4).

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).

Phospho-Actin (Tyr-53) synthetic peptide (coupled to KLH) corresponding to amino acid residues around tyrosine 53 of human β actin*. This sequence is identical to similar regions in all four α actins, as well as in γ actin, and is well conserved in actins from most eukaryotic species.

*For more information, see UniProt Accession P60709

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.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

This antibody detects a 42 kDa* protein corresponding to the molecular mass of Actin on SDS-PAGE immunoblots of pervanadate treated human C2C12 and SYF cSrc transformed cells, but not in control cells. In addition, this antibody either detects actin cleavage products at 36 and 20 kDa, or cross-reacts with unidentified proteins at these molecular weights in C2C12 cells.

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

Product References:

Pryazhnikov, E. et al. (2018) Sci Rep. 8(1):6464. (WB: mouse cortex)
Bertling, E. et al. (2016) J Neurosci. 36(19):5299. (WB/ICC: rat hippocampal neurons)
Vonach, C. et al. (2011) British J Cancer. 105:263. (WB: human endothelial cells)

This kit contains: