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 제품안내  면역학   Cell Structure  Actin Assay

Actin binding protein spin-down Assay Kit, muscle

Actin binding protein은 세포의 형태와 운동성, 근육수축, intracellular trafficking, cell pathogenesis, signal transduction등의 연구에 관여됩니다. Actin binding kit은 고객께서 분리한 단백질의 Actin binding ability를 확인하는데 유용한 kit 입니다.

Cytoskeleton
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특징
  • F-actin 및 actin monomer에 binding하는 단백질을 확인하는데 사용됩니다.
  • muscle tissue로부터 얻은 단백질의 actin filament에 대한 binding 능력을 결정하는데 필요한 시약과 control을 제공합니다.
Kit의 구성
  • 8 x 250 μg Skeletal muscle actin (Cat. # AKL99).
  • α-Actinin, positive control (Cat. # AT01).
  • BSA, negative control
  • General Actin Buffer (Cat. # BSA01).
  • Actin Polymerization Buffer (Cat.# BSA02)
  • F-actin Cushion Buffer.
  • ATP, 100 mM (Cat. no. BSA04)
  • EGTA, 0.5 M
  • MgCl2, 100 mM
  • Tris-HCl pH 6.5, 1.0 M
  • Tris-HCl pH 7.5, 100 mM
  • Manual with detailed protocols and extensive troubleshooting guide.
적용
  • To determine whether a protein binds to filaments or monomers of actin.
  • To determine whether a protein bundles F-actin
  • To test various conditions (e.g. pH optima) or requirements (e.g. divalent cation requirement) for binding to actin.
실험예
The actin binding protein spin-down assay kit was used to study the actin binding and bundling activities of α-actinin (cat. # AT01). α-Actinin was shown to bundle F-actin and make the F-actin sediment at 14,000 x g (Fig. 1).
Figure 1. Actin bundling assay using kit BK001. F-actin was incubated alone or together with α-actinin or BSA. Bundled F-actin was pelleted by a 14,000 x g centrifugation and pellets (P) and supernatants (S) were run on an SDS-PAGE gel. Only in the presence of the F-actin bundling protein α-actinin is actin pelleted at this centrifugation speed.
실험논문
  • Banerjee, J. and Wedegaertner, P. B. (2004). Identification of a novel sequence in PDZ-RhoGEF that mediates interaction with the actin cytoskeleton. Mol. Biol. Cell 15, 1760-1775.
  • Hildebrand, J. D. and Soriano, P. (1999). Shroom, a PDZ domain-containing actin-binding protein, is required for neural tube morphogenesis in mice. Cell 99, 485-497.
  • Kumar, N., Zhao, P., Tomar, A., Galea, C. A. and Khurana, S. (2004). Association of villin with phosphatidylinositol 4,5-bisphosphate regulates the actin cytoskeleton. J. Biol. Chem. 279, 3096-3110.
  • Oliver, C. J., Terry-Lorenzo, R. T., Elliott, E., Bloomer, W. A., Li, S., Brautigan, D. L., Colbran, R. J. and Shenolikar, S. (2002). Targeting protein phosphatase 1 (PP1) to the actin cytoskeleton: the neurabin I/PP1 complex regulates cell morphology. Mol. Cell. Biol. 22, 4690-4701.
  • Takeshita, N., Ohta, A. and Horiuchi, H. (2005). CsmA, a class V chitin synthase with a myosin motor-like domain, is localized through direct interaction with the actin cytoskeleton in Aspergillus nidulans. Mol. Biol. Cell 16, 1961-1970.
  • Torgler, C. N., Narasimha, M., Knox, A. L., Zervas, C. G., Vernon, M. C. and Brown, N. H. (2004). Tensin stabilizes integrin adhesive contacts in Drosophila. Dev. Cell 6, 357-369.
주문정보(Ordering Information)
CATNUM PRODUCT SIZE
BK001Actin Binding Protein Spin-Down Assay Kit30-100 assays