ResourceLife Sciences
Quantitatively Determining the Orientation and Conformation of Proteins in Single Silk Fibers with ATR IR Spectroscopy
27 Jun 2016This application note describes a method using Specac’s Golden Gate ATR Accessory to analyze the orientation and conformation of proteins in silk from the silkworm Bombyx mori. This overall method involves three different methods where the fiber is analyzed from different perspectives.
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Golden Gate® diamond ATR
Specac LtdThe Golden Gate® is a robust option for demanding single-reflection ATR applications. It features a brazed diamond ATR crystal and a signature “Golden Gate” sample loading bridge. More advanced top-plate assemblies are available for in-situ, high- and low-temperature work.
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Atomic Absorption / Emission SpectroscopyAtomic absorption spectroscopy (AAS) and atomic emission spectroscopy (AES) — also called optical emission spectroscopy (OES) — are used to detect the elemental constituents in samples. Both techniques involve the atomization of a sample. Atomic absorption spectrometers may use a flame or furnace to create an atomic vapor of the sample before irradiation with spectral light. Optical emission spectrometers may use a flame, inductively coupled plasma (ICP), microwave plasma (MP) or spark arcs to atomize and excite the sample. At higher excitation energies, electrons can be emitted instead of photons, which can be useful for samples that can’t be atomized and for surface analysis. Explore electron spectroscopy equipment such as Auger spectrometers and photoelectron spectrometers for surface elemental analysis of samples. Find the best atomic absorption, photoelectron and optical emission spectrometers in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Infrared / IR SpectroscopyInfrared (IR) spectroscopy measures the interaction of infrared light with a sample, including transmission, reflectance & absorbance, facilitating the identification of analytes. Equipment used for quantitative analysis includes Fourier-transform infrared (FTIR) spectrometers, infrared cameras, FTIR gas analyzers, as well as attenuated total reflectance (ATR) accessories and pellet or film presses. Find the best IR spectroscopy products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.UV-Visible SpectroscopyUltraviolet-visible (UV-Vis) spectrophotometers are used to measure the interaction of UV and visible light with a sample, including transmission, reflectance & absorbance. The two major instrument classes are single-beam or double-beam spectrophotometers. More specialized equipment includes colorimeters, spectroradiometers and refractometers. Portable and microvolume spectrophotometers are also available. For the modular spectroscopy lab, explore a range of light sources for combination with a spectrograph/spectrometer and optics. Find the best UV-Vis spectroscopy products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.Protein PurificationProtein purification is a vital step in drug discovery, therapeutics, biotech and life science research. The purification process typically involves subcellular or membrane protein extraction with cell lysis kits, separation of proteins from cell debris by filtration or spin columns, and the isolation of proteins of interest from other proteins and impurities with affinity purification (including fusion protein tags and antibody binding proteins A, G and L), immunoprecipitation or chromatographic methods, such as ion exchange, size exclusion and immobilized metal affinity chromatography. All purification methods come in multiple formats for your laboratory needs, including agarose or magnetic beads, resins, columns and filter plates. Find the best protein purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Protein CrystallographyProtein crystallization is the process of crystallizing purified proteins for 3D structure analysis by x-ray crystallography. The main methods of protein crystallization include sitting drop, hanging drop and microbatch. It is important to control parameters such as pH, temperature and concentration. Following crystallization, detectors and software are used for data collection and analysis.NanotechnologyNanotechnology, or nanotech, is an engineering technique using molecular scale functional systems. Applications of nanotechnology include medicine and medical devices, electronics, air and water purification, food science and energy production.Particle CharacterizationParticle characterization instruments are used to determine particle size distribution, shape, surface area, zeta potential, density and porosity of particles and materials. Multiple tecchniques are available for determining particle size, shape and count including dynamic light scattering (DLS), laser diffraction, electrozone (Coulter technique), imaging particle analysis and single particle optical sensing. Determine the density of your material with a gas pycnometer or examine its surface area and porosity with gas adsorption analyzers and mercury porosimeters. Find the best particle characterization instruments in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Non-Destructive TechniquesNon-destructive techniques (NDT) describes a variety of analytical techniques used to evaluate the properties of a material. Common methods include ultrasonic, magnetic-particle, liquid penetrant, radiographic, remote visual inspection (RVI), and eddy-current testing. NDT is regularly used in forensic engineering, civil engineering, mechanical engineering, electrical engineering, systems engineering, aeronautical engineering, and medicine.ProfilometersProfilometers are instruments used to measure a surface's profile, in order to quantify etch depth, deposited film thickness, and surface roughness. They operate in either contact or non-contact modes and may use optical or stylus techniques to make the actual measurements.Electron MicroscopyElectron microscopes (EM) are used to create high-resolution images of samples at the nanoscale by means of an accelerated beam of electrons as a source of illumination. Types of electron microscope include scanning electron microscopes (SEM), transmission electron microscopes (TEM), scanning transmission electron microscopes (STEM) and cryo-electron microscopes. Focused ion beam (FIB) microscopes are useful for modifying or milling a sample surface with nanometer precision, as well as imaging. Find the best electron microscopes in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Protein BiologyThe analysis of protein expression, identity and function is vital for many areas of life science research and drug discovery. Some of the most commonly used techniques in protein analysis include Western blotting, electrophoresis and mass spectrometry.Molecular DesignFTIR PelletsProtein Synthesis