Protein Preparation Protocols

Protein Preparation Protocols

  • Chromatography in Protein purification

    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 […]

  • Desalting and Buffer Exchange with Millipore Centrifugal Devices

  • Methods for Concentrating Protein Solutions

    Describes methods for dialysis, ultrafiltration, ammonium sulfate precipitation (salting out), ion exchange (followed by desalting chromatography), spin-filters, and trichloroacetic acid/deoxycholate (TCA/DOC)

  • Protein Purification (Amersham Handbook)

  • General notes

    Here are a few pieces of information that may be useful to you

  • Designing buffers

    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.

  • Expressing concentration and dilution of stocks

    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.

  • General molecular lab skills and techniques

    By Joni Frederick

  • QIAGEN® Bench Guide

    QIAGEN® Bench Guide, available in PDF

  • BCA assay

    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).

  • Bradford assay

    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.

  • Lowry assay

    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.

  • Nitrilase activity assay

    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.

  • Ultraviolet absorbance of aromatic amino acids

    To different degrees, all aromatic amino acids absorb ultraviolet light.

  • Introduction to general crystallography

    Crystallography is the experimental science of determining the arrangement of atoms in the crystalline solids.

  • Agarose gel electroporesis

    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.

  • SDS-PAGE Technique

    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.

  • SDS-PAGE gel staining

    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.

  • High Sensitivity Silver Stain for SDS-PAGE

    Sigma-Aldrich protocol for ProteoSilver™: High Sensitivity Silver Stain for SDS-PAGE

  • Imidazole–SDS–zinc reverse staining

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