Acrylodan, reacted with thiols, is sensitive to the local environmental dipolarity and dynamics within the binding pocket surrounding Cys^34[1][2].

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.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

225.29

C₁₅H₁₅NO

86636-92-2

Room temperature in continental US; may vary elsewhere.

Powder	-20deg;C	3 years
	4deg;C	2 years
In solvent	-80deg;C	6 months
	-20deg;C	1 month

In Vitro:;

DMSO : 50 mg/mL (221.94 mM; Need ultrasonic)

*

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°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

• [1]. Sarah F Stuart, et al. A two-step mechanism for the folding of actin by the yeast cytosolic chaperonin. J Biol Chem. 2011 Jan 7;286(1):178-84.

  [2]. K Flora, et al. Unfolding of acrylodan-labeled human serum albumin probed by steady-state and time-resolved fluorescence methods. Biophys J. 1998 Aug;75(2):1084-96.