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Acrylodan

CAS No. : 86636-92-2

MCE 站：Acrylodan

产品活性：Acrylodan 可与硫醇反应，对 Cys³⁴ 周围的结合口袋内的局部环境双极性和动力学敏感。

研究领域：Others

作用靶点：Others

In Vitro: Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Purification and Labeling of Act1:
1.20-fold molar excess of acrylodan (Anaspec) was added to the polymerized actin and incubated on ice overnight.
2.The labeled actin is centrifuged (2 h, 4 °C, 100,000 × g) then the pellet was resuspended in 10 mm Tris, pH 7.5, 0.5 mm β-mercaptoethanol, 0.2 mm CaCl2, 50 μm ATP and dialyzed against the same buffer for 70 h at 4 °C.
3.The supernatant was collected and the actin concentration and degree of labeling were calculated using the extinction coefficient 26,600 M^?1 cm^?1 for actin at 290 nm, 18,500 M^?1 cm^?1 or acrylodan at 385 nm, and 71,000 M^?1 cm^?1 for Alexa Fluor 488 at 494 nm.
4.Acrylodan-labeled Act1 (^AcrylAct1_NAT) is dramatically different than that of EDTA-unfolded ^AcrylAct1 (^AcrylAct1I3), which has a very similar emission spectrum to CCT-bound ^AcrylAct1.

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