Most of the information you'll ever need is provided on the Amersham website (under Literature » Handbooks). It covers basic principles of protein purification, basic theory of chromatography, how to pack columns, assess packing efficiency, recommended flow rates etc. Other documents on these topics are: One important consideration in gel filtration chromatography is to always […]
Describes methods for dialysis, ultrafiltration, ammonium sulfate precipitation (salting out), ion exchange (followed by desalting chromatography), spin-filters, and trichloroacetic acid/deoxycholate (TCA/DOC)
Here are a few pieces of information that may be useful to you
A buffer is a mixture of dissolved substances which tends to keep the pH of a solution constant when modest additions of acid and base are made to that solution.
Concentration of a solution represents the amount of solute dissolved in a given amount of solvent or solution. It is a macroscopic property and can be expressed in a variety of ways; qualitatively and quantitatively.
By Joni Frederick
QIAGEN® Bench Guide, available in PDF
The bicinchoninic acid assay (BCA assay), also known as the Smith assay, is a biochemical assay for determining the total concentration of protein in a solution (0.5 μg/mL to 1.5 mg/mL).
The Bradford protein assay is a spectroscopic analytical procedure used to measure the concentration of protein in a solution. It is subjective, i.e., dependent on the amino acid composition of the measured protein.
The Lowry protein assay is a biochemical assay for determining the total level of protein in a solution. The total protein concentration is exhibited by a color change of the sample solution in proportion to protein concentration, which can then be measured using colorimetric techniques.
1. Picric acid colorimetric assay for the quantitative determination of cyanide - for activity assays of cyanide-degrading nitrilases.
2. Indophenol blue method for the quantitative detection of ammonia/ammonium - for activity assays of nitrilases that hydrolyze higher nitriles (benzonitrile among others) resulting in the production of ammonia among other products.
To different degrees, all aromatic amino acids absorb ultraviolet light.
Crystallography is the experimental science of determining the arrangement of atoms in the crystalline solids.
Agarose gel electrophoresis separates DNA fragments according to their size. Typically, a DNA molecule is digested with restriction enzymes, and the agarose gel electrophoresis is used as a diagnostic tool to visualize the fragments. An electric current is used to move the DNA molecules across an agarose gel, which is a polysaccharide matrix that functions as a sort of sieve to help "catch" the molecules as they are transported by the electric current.
For proteins, sodium dodecyl sulfate (SDS) is an anionic detergent applied to protein sample to linearize proteins and to impart a negative charge to linearized proteins. This procedure is called SDS-PAGE.
Once protein bands have been separated by electrophoresis, they can be visualized using different methods of in-gel detection. The choice of staining technique depends on the availability of imaging equipment in the lab in many cases.
Sigma-Aldrich protocol for ProteoSilver™: High Sensitivity Silver Stain for SDS-PAGE