86636-92-2
基本信息
6-丙烯?;?2-二甲氨基萘
1-(6-(二甲基氨基)萘-2-基)丙-2-烯-1-酮
6-ACRYLOYL-2-DIMETHYLAMINONAPHTHALENE
1-(6-Dimethylamino-naphthalen-2-yl)-propenone
Acrylodan [6-Acryloyl-2-dimethylaminophthalene]
ACRYLODAN [6-ACRYLOYL-2-DIMETHYLAMINONAPHTHALENE]
1-(6-(Dimethylamino)-2-naphthalenyl)-2-propen-1-one
1-(6-(Dimethylamino)naphthalen-2-yl)prop-2-en-1-one
2-Propen-1-one, 1-(6-(dimethylamino)-2-naphthalenyl)-
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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 (
Acryl
Act1
NAT
) is dramatically different than that of EDTA-unfolded
Acryl
Act1 (
Acryl
Act1I3), which has a very similar emission spectrum to CCT-bound
Acryl
Act1.