F-actin visualization Assay Kit

• 일반 많은 actin 항체와 같이 monomeric G-actin에 결합하지 않기 때문에 깔끔한 filament staining 결과를 얻을 수 있습니다.
• Non-specific staining이 거의 없습니다.

제품의 구성
• Rhodamine Phalloidin (Cat. # PHDR1)
• Fixation Buffer
• Permeabilization Buffer
• Wash buffer
• Mounting Medium
• Dark Box
• Manual with detailed protocols and extensive troubleshooting guide
• staining 300 coverslips (12 mm circular)

• Cellular colocalization with actin binding proteins by immunofluorescence.
• Detecting changes in actin morphology upon bacterial or viral infections.
• Detecting changes in actin morphology upon activation of small G-protein signal transduction pathways (see Fig. 1)
• Focal adhesion marker
• Marker for serum starvation of tissue culture cells (see Fig. 1)

실험예
• Figure 1. below shows 3T3 cells under serum starvation conditions (A) and calpeptin stimulated conditions (B) stained with the F-actin Visualization Biochem Kit. It can be seen that most actin stress fibers disappear under serum starvation conditions, due to the de-activation of the Rho signaling pathway, while calpeptin stimulation of the cells results in a rapid accumulation of actin stress fibers due to the activation of Rho (4).

Figure 1. Phalloidin stained serum starved and calpeptin treated cells. Swiss 3T3 cells were grown in DMEM plus 10% calf bovine serum to 60% confluency, mediium was changed to 0.5% calf bovine serum for 24 h then to 0% calf bovine serum for an additional 24 h. After this serum starvation procedure, one dish was treated with calpeptin (0.1 mg/ml final) for 30 min while the other dish treated with carrier only (DMSO). After incubation, the coverslips were removed from the dishes and stained according to the BK005 manual. A: serum starved cells, B: calpeptin treated cells).

Product Citations
• Sheterline, P., Clayton, J., & Sparrow, J.C (Eds.). (1998) Protein Profile: Actin. 4th edition. Oxford University Press.
• dos Remedios, C.G. et al. (2003) Actin binding proteins: Regulation of cytoskeletal microfilaments. Physiol. Rev. 83: 433-473.
• Maciver S.K. The encyclopaedia of actin-binding proteins (and drugs)
• Ridley, A. J. and Hall, A. (1992) The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell 70: 389-399 (1992).
• Merz, A. J. and Higgs, H. N. (2003) Listeria Motility: Biophysics Pushes Things Forward. Curr. Biol. 13: R302-R304.
• Wulf, E., et al. (1979) Fluorescent phallotoxin: a tool for the visualization of cellular actin. Proc. Natl. Acad. Sci. USA 76: 4498-4502.