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Technical Data Sheet

AX1675

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phospho-Actin (Tyr-53)
Blocking Peptide
Price
Size

$55
50 μg

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Enlarge

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).

Application
Dilution


Blocking
1:1000


ELISA
50 ng/well



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
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 organisation 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 polymerizaiton. In B cells, SHP-1 tyrosine dephosphorylation of actin leads to actin filament depolymerization following BCR stimulation.


Background References
Jungbluth, A. et al. (1995) FEBS Let. 375:87.
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.
Sequence
Actin synthetic peptide corresponding to amino acid residues in the N-terminal region 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.
Buffer and Storage
Blocking Peptide is supplied in 50µl phosphate-buffered saline and 0.05% sodium azide.
Store at –20°C. Stable for 1 year.
Specificity
The peptide is specifically recognized by actin (Tyr-53) phospho-specific antibody (AP1671) in ELISA, and has been shown to block the reactivity of AP1671 in Western blot and immunocytochemistry.
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