Skip to content

JASCO JASCO

  • News
  • Events
  • E-Store
  • My Account
  • Contact Us
  • Worldwide
Search
Click to view menu
MENUMENU
  • Products
    • Chromatography
      • HPLC
      • RHPLC
      • UHPLC
      • LC-MS
      • Preparative LC
      • Analytical SFC
      • Semi-Preparative SFC
      • Hybrid SFC
      • Fuel Analysis by SFC-FID
      • Preparative SFC
      • Supercritical Fluid Extraction
      • Chromatography Software
    • Molecular Spectroscopy
      • Circular Dichroism
      • High-Throughput CD
      • Vibrational CD
      • Circularly Polarized Luminescence (CPL)
      • Polarimeters
      • FTIR Spectrometers
      • FTIR Microscopy
      • FTIR Portable
      • Raman Microscopy
      • Palmtop Raman Spectrometer
      • Probe Raman
      • UV-Visible/NIR Spectrophotometers
      • UV-Visible/NIR Microscopy
      • Fluorescence Spectrophotometers
      • Film Thickness
      • Spectra Manager™ Suite
    • Refurbished
      • Refurbished HPLC Systems
      • HPLC Switching Valves
      • NRS-4100 Confocal Raman Microscopes
      • FTIR Accessories
  • Service
    • Service and Support Plans
    • Service Request Form
  • Applications
  • KnowledgeBase
  • Learning Center
    • Best Practices
      • Circular Dichroism Tips & Tricks for Biological Samples
      • CD Scale Calibration with ACS
      • Fluorescence Tips & Tricks
      • Raman Spectroscopy Tips & Tricks
    • Training Videos
      • ChromNAV
      • SF-NAV
      • Circular Dichroism
      • UV-Visible/NIR
      • Fluorescence
    • Training Seminars
      • Registration Form
    • Webinars
    • eBooks
    • Theory
      • Theory of Molecular Spectroscopy
      • Chromatography
  • About Us
    • President’s Message
    • Contact
    • History
    • Careers
  • News
  • Events
  • Worldwide
  • Shop
  • My Account

Home / Applications / CPL Spectrum Measurement of Green Fluorescent Protein (GFP)

  • Industry

  • Technique

CPL Spectrum Measurement of Green Fluorescent Protein (GFP)

By Leah Pandiscia, PhD

image_pdfView and Download PDF
August 15, 2022

Introduction

Circularly polarized luminescence Spectrophotometer
CPL-300 Circularly Polarized Luminescence Spectrophotometer

Circularly polarized luminescence (CPL) measures the difference in the emission intensities of left- and right-handed circularly polarized light of chiral compounds. Unlike CD, CPL provides information on the excited state properties of chiral molecules.

Green fluorescence protein (GFP) is a protein that emits green fluorescence when exposed to ultraviolet light and has been shown to have chiroptical properties1. This application note illustrates the CPL and CD spectra of GFP obtained with the CPL-300 and J-1500, respectively.

Experimental

Measurement Conditions
CPL-300 Circular Polarized Luminescence
Excitation Wavelength399 nm
Excitation Bandwidth
12 nm
Data Interval
1 nm
Response
16 seconds
Emission Bandwidth
8 nm
Scan Speed
10 nm/min
Accumulations
36
Path length
10 mm
Measurement Conditions
J-1500 Circular Dichroism Spectrophotometer
Data Pitch0.2 nm
D.I.T.1 sec
Bandwidth1 nmScan Speed200 nm/min
Accumulations16Path Length20 mm

Keywords

J-1500, Circular dichroism, CPL-300, Circularly polarized luminescence, Protein structure, Fluorescence, Biochemistry

Results

CD and absorption spectra (Figure 1, green) of a 0.03 mg/mL GFP solution was measured in the near-UV and visible regions. The GFP chromophore is observed in the visible region at 400 nm and the absorption bands due to the aromatic amino acid residues are observed in the near-UV region between 250 and 300 nm.  The CPL and fluorescence spectra were also obtained and show in blue in Figure 1. The characteristic fluorescent and CPL spectra of GFP are observed between 470-570 nm.

CPL (blue) and CD (green) spectra (top) of green fluorescent protein. The absorption (green) and fluorescence (blue) spectra are shown at the bottom.
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.

image_pdfView and Download PDF

Featured Products:

  • Circularly Polarized Luminescence

  • J-1700 Circular Dichroism Spectrophotometer

    UV, Visible, and NIR measurements up to 2500 nm for MCD and specialized applications.

    J-1700

  • VCD Icon Bottom Aligned

    VCD Spectrometer

About the Author

Leah Pandiscia received her PhD from Drexel University where she studied Biophysical Chemistry. She is a Spectroscopy Applications Scientist.

JASCO provides services and support to a diverse range of industries throughout the world.

View our support plans

Connect with JASCO

  • Facebook
  • Twitter
  • LinkedIn
  • JASCO Sales
  • 800-333-5272

Receive the latest promotions and special offers

  • Careers
  • Press Kit
  • Privacy Policy
  • Sitemap

© , JASCO. All Rights Reserved.