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Quantitation of the metal content of proteins by total reflection X-ray fluorescence (TXRF) spectrometry (CAT#: STEM-ST-0201-WXH)

Introduction

It is estimated that approximately half of all proteins contain a metal. In another estimate, about one quarter to one third of all proteins are proposed to require metals to carry out their functions. Thus, metalloproteins have many different functions in cells, such as storage and transport of proteins, enzymes and signal transduction proteins, or infectious diseases. The abundance of metal binding proteins may be inherent to the amino acids that proteins use, as even artificial proteins without evolutionary history will readily bind metals.




Principle

XRF describes the process where some high-energy radiation excites atoms by shooting out electrons from the innermost orbitals. When the atom relaxes, that is, when outer electrons fill inner shells, X-Ray fluorescence radiation is emitted.
Total-reflection X-ray fluorescence (TXRF) is a multi-element simultaneous analysis technology developed based on X-ray fluorescence (XRF). An aircooled X-ray tube generates an X-ray beam, which is reduced to a narrow energy range by a multi-layer monochromator. The fine beam impinges on a polished sample carrier at a very small angle and is totally reflected.

Applications

XRF is widely used as a fast characterization tool in many analytical labs across the world, for applications as diverse as metallurgy, forensics, polymers, electronics, archaeology, environmental analysis, geology and mining.

Procedure

1. Primary X-rays knock out an electron from one of the orbitals surrounding the nucleus within an atom of the material.
2. A hole is produced in the orbital, resulting in a high energy, unstable configuration for the atom.
3. To restore equilibrium, an electron from a higher energy, outer orbital falls into the hole. Since this is a lower energy position, the excess energy is emitted in the form of fluorescent X-rays.
The energy difference between the expelled and replacement electrons is characteristic of the element atom in which the fluorescence process is occurring – thus, the energy of the emitted fluorescent X-ray is directly linked to a specific element being analyzed.

Materials

XRF spectrometer (including X-ray source, sample chamber, analysing crystal, detector and signal processing computer)
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