Lecture 4. Spectrophotometry of Biomolecules Flashcards
What is spectroscopy ?
An experimental technique for monitoring the interaction of electromagnetic radiation with matter by measuring changes in intesity of electromagnetic radiation
What is a spectrum representes as ?
Intensuty verses energy
What is there an inverse relationship between ?
Wavelength and energy
What are the two types of spectroscopy ?
- Absorption spectroscopy
- Emission spectroscopy
What is the basic protocol for absorption spectroscopy ?
Irradiate the sample at a specific frequency and measure the amount of radiation absorbed
What is the basic protocol for emission spectroscopy ?
Irradiate the sample and measure the intensity of the emitted radiation at a specific frequency
What does electromagnetic radiation consist of ?
An oscillating electric field and an oscillating magnetic field at right angles
What can an electromagnetic radiation induce ?
Oscillations in any species that contains an electric or magnetic dipole
What are the properties of gamma rays ?
Very penetratinf can ionise molecules leading to chemical changes
What are the properties of Near UV infared rays ?
Not very penetrating, interation with molecules produces heat
What are the properties of micro/radio waves ?
Inteact weakly with molecules
What is a chiral molecule ?
One that is not superimposable on its mirror image
What is the intensity of fluorescence emission directly proportional to ?
Intensity of the incident light
What does the sensitivity of the instrument depends on ?
- The nature of the light source
- The slit width used
- Electronic amplification system
What will the sensititvity also depend on ?
- Concentration
- Temperature
- Molar extinction coefficient
What are the reporter groups involved in applying fluorescence spectroscopy to proteins ?
- Intrinsic fluorophores
- Extrinsic fluorophores
