Acrylodan 

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.

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

225.29

C₁₅H₁₅NO

86636-92-2

Solid

Light yellow to yellow

C=CC(C1=CC=C2C=C(C=CC2=C1)N(C)C)=O

Room temperature in continental US; may vary elsewhere.

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [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.  [Content Brief]

  [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.  [Content Brief]