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Technical Data Sheet

AK7600

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Actin Filament Regulation
Immunocytochemistry Kit
Price

$355

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
Kit Summary
The actin filament regualtion kit can be used to compare labeling between actin filaments and total actin. The kit contains two antibodies for detecting total actin, and Phalloidin:FITC for labeling actin filaments. In addition, secondary reagents conjugated to DyLight® 594 are included for dual labeling experiments with Phalloidin:FITC.
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Immunocytochemical labeling of actin filaments in A7r5 cells. Methanol and acetone fixed cells were labeled with mouse monoclonal anti-Actin (AM2021) and rabbit polyclonal anti-Actin (AP1651). The antibodies were detected using Goat anti-Mouse or Goat anti-Rabbit DyLight® 594. Paraformaldehyde-fixed and NP-40-permeabilized cells were labeled with Phalloidin:FITC.

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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
Wulf, E. et al. (1979) Proc. Nat. Acad. of Sci.(USA). 76:4498.
Faulstich, H. et al. (1983) Exp. Cell Res. 144:73.
Faulstich, H. et al. (1988) J. Muscle Res. Cell Motility. 9:370.
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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