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Technical Data Sheet

AP1651

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Actin (N-terminal region)
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$245
100 μl
Hu, Rt, Ms, Ck
42 kDa

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Enlarge

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).

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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).

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Application
Dilution


ELISA
1:2000


ICC
1:50


IHC
1:100


IP
1:25


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 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 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.


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.
Immunogen
Actin synthetic peptide (coupled to KLH) 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
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.
Specificity
This antibody detects a 42 kDa* protein corresponding to the molecular mass of Actin on SDS-PAGE immunoblots of human A431, SYF, and HUVEC cells, as well as mouse 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.
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