绿色荧光蛋白中手性检测方案(圆二色光谱仪)

Application Note 2/2 CD-0030 CPL Spectrum Measurement of Green FluorescentProtein (GFP) Introduction Circularly polarized luminescence (CPL) measures thedifference in the emission intensities of left- and right-handedcircularly polarized light of chiral compounds. Unlike CD， CPLprovides information on the excited state properties of chiralmolecules. CPL-300 Circularly Polarized Luminescence Spectrophotometer Green fluorescence protein (GFP) is a protein that emits green fluorescence when exposed to ultraviolet light and has been shown to have chiroptical properties. This application noteillustrates the CPL and CD spectra of GFP obtained with the CPL-300 and J-1500， respectively. Keywords J-1500， Circular dichroism， CPL-300， Circularly polarized luminescence， Protein structure， Fluorescence， Biochemistry Experimental Measurement Conditions CPL-300 Circularly Polarized Luminescence Excitation Wavelength 399 nm Excitation Bandwidth 12 nm Data Interval 1nm Response 16 seconds Emission Bandwidth 8nm Scan Speed 10 nm/min Accumulations 36 Path Length 10 mm Measurement Conditions J-1500 Circular Dichroism Spectrophotometer Data Pitch 0.2 nm D.I.T. 1 sec Bandwidth 1nm Scan Speed 200 nm/min Accumulations 16 Path Length 20mm CPL Spectrum Measurement ofApplication Note Green Fluorescent Protein (GFP) Results CD and absorption spectra (Figure 1， green) of a 0.03 mg/mL GFP solution was measured in the near-UV and visibleregions. The GFP chromophore is observed in the visible region at 400 nm and the absorption bands due to the aromaticamino acid residues are observed in the near-UV region between 250 and 300 nm. The CPL and fluorescence spectrawere also obtained and show in blue in Figure 1. The characteristic fluorescent and CPL spectra of GFP are observedbetween 470-570 nm. Figure 1. CPL (blue) and CD (green) spectra (top) of green fluorescent protein. The absorption (green) and fluorescence (blue)spectra are shown at the bottom. References 1. Hiromasa Goto， Isao Sawada， and Nobuhiko Nomura， International Journal of Polymeric Materials， 2010， 59， 786-792. JASCO INC. 28600 Mary's Court， Easton， MD 21601 USA JASCO INC.Mary's Court， Easton， MD USATel： ( Fax： (pplication Library： http：//www.jascoinc.com/applications Tel： ( Fax： ( pplication Library： http：//www.jascoinc.com/applicationsCopyright O JASCO Corporation 圆偏振荧光光谱仪(CPL)测量手性化合物的左、右圆偏振光发射强度的差异。与圆二色光谱仪（CD）不同，CPL提供手性分子激发态性质信息。