4-Di-10-ASP is a fluorescent lipophilic tracer (Excitation 485 nm; Emission 620 nm). 4-Di-10-ASP can be used to stain phospholipid membranes in a specific manner[1][2].
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1. 4-Di-10-ASP (1 μM), DOPC (10 mM), and DOPG (1 mM) is dissolved in methanol/chloroform (25 L, 1:2 v/v).
2. The solution is allowed to dry overnight under vacuum to obtain lamellar lipid films, which in turn are hydrated with the transcription/translation solution (25 L) for three hours at 37℃.
3. An aliquot (10 L) of the solution thus prepared is placed on a glass slide and sealed by a cover glass.
4. The sample is immediately observed with a confocal laser-scanning microscope, an argon laser (488 nm) is employed to excite the 4-Di-10-ASP[2].
[1]. Z J Huang, et al. Partition coefficients of fluorescent probes with phospholipid membranes. Biochem Biophys Res Commun. 1991 Nov 27;181(1):166-71.
[2]. Shin-ichiro M Nomura, et al. Gene expression within cell-sized lipid vesicles. Chembiochem. 2003 Nov 7;4(11):1172-5.