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The Photoreactions of Recombinant Phytochrome by UV-Vis Spectroscopy (CAT#: STEM-MB-0998-WXH)

Introduction

In photomorphogenesis of green plants phytochromes play a decisive role in perception of light and in regulating numerous responses of the plants to optimize their growth and development with respect to the light conditions of the environment. The chromoproteins are characterized by an interconvertible photoreaction between two parent states, i.e. the red-absorbing form and the physiologically active, far-red–absorbing form.
The transduction of the light-generated signal and the identification of the signal transduction pathway are still a major field in phytochrome research. While for plants a variety of pathways and participating components are discussed, phytochromes and phytochrome-like proteins in prokaryotes are thought to act as light-regulated histidine protein kinases.




Principle

UV-Vis spectroscopy is an analytical technique that measures the amount of discrete wavelengths of UV or visible light that are absorbed by or transmitted through a sample in comparison to a reference or blank sample. This property is influenced by the sample composition, potentially providing information on what is in the sample and at what concentration. The only requirement is that the sample absorb in the UV-Vis region, i.e. be a chromophore. Absorption spectroscopy is complementary to fluorescence spectroscopy. Parameters of interest, besides the wavelength of measurement, are absorbance (A) or transmittance (%T) or reflectance (%R), and its change with time.

Applications

UV/Vis spectroscopy is routinely used in analytical chemistry for the quantitative determination of diverse analytes or sample, such as transition metal ions, highly conjugated organic compounds, and biological macromolecules. Spectroscopic analysis is commonly carried out in solutions but solids and gases may also be studied.

Procedure

1. Calibrate the Spectrometer
2. Perform an Absorbance Spectrum
3. Kinetics Experiments with UV-Vis Spectroscopy

Materials

UV/VIS Spectrophotometer
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