G-actin : F-actin in vivo Assay Kit
F-actin 대 G-actin의 양을 비교측정하기 위해서 western blot 방법이 이용되며, 이 kit은 western blot 실험에 필요한 시약이 제공됩니다.
- G-actin pool로부터 F-actin을 원심분리로 얻은뒤, SDS-PAGE와 Western blot으로 그양을 측정합니다.
- 세포질내에 있는 G-actin 대 세포골격내에 있는 F-actin의 ratio를 비교할수 있습니다.
- Lysis and F-actin stabilization buffer
- ATP (Cat. # BSA04)
- Protease inhibitor cocktail
- F-actin enhancing control solution
- F-actin depolymerization control solution
- Control G-actin Standard (Cat. # AKL99)
- Actin polyclonal antibody (Cat. # AAN01)
- SDS sample buffer (5 x)
- Manual with detailed protocols and extensive troubleshooting guide
- To study the effects of pharmaceutical compounds on the ratio of G-actin to F-actin.
- To study the effects of mutated cell lines versus their parent cell line for the change in ratio of G-actin to F-actin.
- To study the effects of physical alterations of environment on the ratio of G-actin to F-actin.
Changes in the amount of G-actin and F-actin were investigated in Swiss 3T3 cells treated with the actin polymerizing drug jasplakinolide, using the G-actin/F-actin in vivo assay kit. In untreated Swiss 3T3 cells, 80% of actin is soluble G-actin, and is found within the supernatant fraction, 20% of actin is filamentous F-actin and is found in the pellet fraction. In Swiss 3T3 cells treated with jasplakinolide, 80% of actin is reorganized into F-actin and is found in the pellet fraction (Fig. 1).
Figure 1. Actin polymerization stimulated by Arp2/3 complex and the VCA domain of WASP. Actin polymerization was measured using kit BK003. The addition of Arp2/3 complex or the VCA domain alone to actin has minimal effects on actin polymerization, while the combination of Arp2/3 and the VCA domain strongly stimulates the rate of actin polymerization.
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- Tang, D. D. and Gunst, S. J. (2004). The small GTPase Cdc42 regulates actin polymerization and tension development during contractile stimulation of smooth muscle. J. Biol. Chem. 279, 51722-51728.
- Tu, Y., Wu, S., Shi, X., Chen, K. and Wu, C. (2003). Migfilin and Mig-2 link focal adhesions to filamin and the actin cytoskeleton and function in cell shape modulation. Cell 113, 37-47.
- Zhang, W., Wu, Y., Du, L., Tang, D. D. and Gunst, S. J. (2005). Activation of the Arp2/3 complex by N-WASp is required for actin polymerization and contraction in smooth muscle. Am. J. Physiol. 288, C1145-1160.