Acrylodan |
Katalog-Nr.GC61505 |
Mit Thiolen umgesetztes Acrylodan reagiert empfindlich auf die lokale DipolaritÄt und Dynamik der Umgebung innerhalb der Bindungstasche, die Cys34 umgibt.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 86636-92-2
Sample solution is provided at 25 µL, 10mM.
Acrylodan, reacted with thiols, is sensitive to the local environmental dipolarity and dynamics within the binding pocket surrounding Cys34[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 (AcrylAct1NAT) is dramatically different than that of EDTA-unfolded AcrylAct1 (AcrylAct1I3), which has a very similar emission spectrum to CCT-bound AcrylAct1.
[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.
Average Rating: 5
(Based on Reviews and 3 reference(s) in Google Scholar.)GLPBIO products are for RESEARCH USE ONLY. Please make sure your review or question is research based.
Required fields are marked with *