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 제품안내  면역학   Cell Structure  Actin Assay

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.

Cytoskeleton
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특징
  • 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.
Kit의 구성
  • 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)
  • DMSO
  • Manual with detailed protocols and extensive troubleshooting guide
적용
  • 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.
실험예
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.
실험논문
  • Chen, G., Raman, P., Bhonagiri, P., Strawbridge, A. B., Pattar, G. R. and Elmendorf, J. S. (2004). Protective effect of phosphatidylinositol 4,5-bisphosphate against cortical filamentous actin loss and insulin resistance induced by sustained exposure of 3T3-L1 adipocytes to insulin. J. Biol. Chem. 279, 39705-39709.
  • Meeks, M. K., Ripley, M. L., Jin, Z. and Rembold, C. M. (2005). Heat shock protein 20-mediated force suppression in forskolin-relaxed swine carotid artery. Am. J. Physiol. 288, C633-639.
  • 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.
주문정보(Ordering Information)
CATNUM PRODUCT SIZE
BK037G-actin / F-actin in vivo assay kit30-100 assays