Search results
Results from the WOW.Com Content Network
Cycloheximide chases are also valuable for assessing how different mutations affect the stability of a protein. Experiments have been conducted in yeast and mammalian cells to determine the critical residues required for protein stability and how disease-associated mutations may be affecting protein half-lives within the cell.
Size exclusion chromatography can be used directly to access protein stability in the presence or absence of ligands. [31] Samples of purified protein are heated in a water bath or thermocycler, cooled, centrifuged to remove aggregated proteins, and run on an analytical HPLC. As the melting temperature is reached and protein precipitates or ...
SILAC has emerged as a very powerful method to study cell signaling, post translation modifications such as phosphorylation, [6] [7] protein–protein interaction and regulation of gene expression. In addition, SILAC has become an important method in secretomics, the global study of secreted proteins and secretory pathways. [8]
In molecular biology, alanine scanning is a site-directed mutagenesis technique used to determine the contribution of a specific residue to the stability or function of a given protein. [1] Alanine is used because of its non-bulky, chemically inert, methyl functional group that nevertheless mimics the secondary structure preferences that many ...
The Instability index is a measure of proteins, used to determine whether it will be stable in a test tube. If the index is less than 40, then it is probably stable in the test tube. If it is greater (for example, enaptin ) then it is probably not stable.
Baculovirus-infected insect cells [20] (Sf9, Sf21, High Five strains) or mammalian cells [21] (HeLa, HEK 293) allow production of glycosylated or membrane proteins that cannot be produced using fungal or bacterial systems. [20] [6] It is useful for production of proteins in high quantity. Genes are not expressed continuously because infected ...
The reagents in this method tend to stain the test tubes. Same test tubes cannot be used since the stain would affect the absorbance reading. This method is also time sensitive. When more than one solution is tested, it is important to make sure every sample is incubated for the same amount of time for accurate comparison. [18]
Ammonium sulfate is an inorganic salt with a high solubility that disassociates into ammonium (NH + 4) and sulfate (SO 2− 4) in aqueous solutions. [1] Ammonium sulfate is especially useful as a precipitant because it is highly soluble, stabilizes protein structure, has a relatively low density, is readily available, and is relatively inexpensive.