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In the less extensive technique of equilibrium unfolding, the fractions of folded and unfolded molecules (denoted as and , respectively) are measured as the solution conditions are gradually changed from those favoring the native state to those favoring the unfolded state, e.g., by adding a denaturant such as guanidinium hydrochloride or urea.
Buffer capacity rises to a local maximum at pH = pK a. The height of this peak depends on the value of pK a. Buffer capacity is negligible when the concentration [HA] of buffering agent is very small and increases with increasing concentration of the buffering agent. [3] Some authors show only this region in graphs of buffer capacity. [2]
In biochemistry, denaturation is a process in which proteins or nucleic acids lose folded structure present in their native state due to various factors, including application of some external stress or compound, such as a strong acid or base, a concentrated inorganic salt, an organic solvent (e.g., alcohol or chloroform), agitation and radiation, or heat. [3]
A thermal shift assay (TSA) measures changes in the thermal denaturation temperature and hence stability of a protein under varying conditions such as variations in drug concentration, buffer formulation (pH or ionic strength), redox potential, or sequence mutation. The most common method for measuring protein thermal shifts is differential ...
Many research and clinical examples [5] exist in the literature that show the use of melting curve analysis to obviate or complement sequencing efforts, and thus reduce costs. While most quantitative PCR machines have the option of melting curve generation and analysis, the level of analysis and software support varies.
Note that the coefficients in front of the "x" correlate to the mole ratios of the reactants to the product. For example, if the reaction equation had 2 H + ions in the product, then the "change" for that cell would be "2x" The fourth row, labeled E, is the sum of the first two rows and shows the final concentrations of each species at equilibrium.
C 0 t analysis, a technique based on the principles of DNA reassociation kinetics, is a biochemical technique that measures how much repetitive DNA is in a DNA sample such as a genome. [1] It is used to study genome structure and organization and has also been used to simplify the sequencing of genomes that contain large amounts of repetitive ...
Radioimmunoprecipitation assay buffer (RIPA buffer) is a lysis buffer used to lyse cells and tissue for the radio immunoprecipitation assay (RIPA). [1] [2] This buffer is more denaturing than NP-40 or Triton X-100 because it contains the ionic detergents SDS and sodium deoxycholate as active constituents and is particularly useful for disruption of nuclear membranes in the preparation of ...