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Technical Data Sheet

AK7710

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Actin Phospho-Regulation
Immunocytochemistry Kit
Price

$405

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
AP1671 Vonach, C. et al. (2011) British J Cancer. 105:263. WB: human endothelial cells
Kit Summary
The actin phospho-regualtion kit can be used to label actin phosphorylation at Tyr-53 relative to total actin expression pattern. The kit contains anti-Actin (Tyr-53) phospho-specific antibody, and two antibodies for detecting total actin. In addition, secondary reagents conjugated to DyLight® 488 or DyLight® 594 are included for labeling primary antibodies for detection using green (493Ex/518Em) or red (593Ex/618Em) filter sets.
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Immunocytochemical labeling of phosphorylated actin in paraformaldehyde fixed and acetone permeabilized C2C12 cells treated with pervanadate. The cells were labeled with mouse monoclonal anti-Actin (AM2021) and rabbit polyclonal anti-Actin (AP1651) or anti-Actin (Tyr-53) (AP1671). The antibodies were detected using Goat anti-Mouse DyLight® 594 or Goat anti-Rabbit DyLight® 488.

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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.
Buffer and Storage
Mouse monoclonal, rabbit polyclonal, and secondary reagents are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
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