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。
| 生物活性 | Acrylodan, reacted with thiols, is sensitive to the local environmental dipolarity and dynamics within the binding pocket surrounding Cys34[1][2]. |
体外研究
(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 ℃, 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 ℃.
3. The supernatant was collected and the actin concentration and degree of labeling were calculated using the extinction coefficient 26,600 M–1cm–1for actin at 290 nm, 18,500 M–1cm–1or acrylodan at 385 nm, and 71,000 M–1cm–1for Alexa Fluor 488 at 494 nm.
4. Acrylodan-labeled Act1 (AcrylAct1NAT) is dramatically different than that of EDTA-unfoldedAcrylAct1 (AcrylAct1I3), which has a very similar emission spectrum to CCT-boundAcrylAct1.
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| 分子量 | |
| 性状 | |
| Formula | |
| CAS 号 | |
| Excitation (Ex) | Ex<380 nm Ultraviolet; Ex 450-495 nm Blue |
| 运输条件 | Room temperature in continental US; may vary elsewhere. |
| 储存方式 | | Powder | -20°C | 3 years | | 4°C | 2 years | | In solvent | -80°C | 6 months | | -20°C | 1 month |
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| 溶解性数据 | In Vitro: DMSO : 50 mg/mL(221.94 mM;Need ultrasonic) 配制储备液 | 1 mM | 4.4387 mL | 22.1936 mL | 44.3872 mL | | 5 mM | 0.8877 mL | 4.4387 mL | 8.8774 mL | | 10 mM | 0.4439 mL | 2.2194 mL | 4.4387 mL |
*请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80℃, 6 months; -20℃, 1 month。-80℃ 储存时，请在 6 个月内使用，-20℃ 储存时，请在 1 个月内使用。 |
