SDS-PAGE

related topics
{acid, form, water}
{math, energy, light}
{rate, high, increase}
{line, north, south}
{system, computer, user}
{area, part, region}
{album, band, music}
{style, bgcolor, rowspan}

SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis, is a technique widely used in biochemistry, forensics, genetics and molecular biology to separate proteins according to their electrophoretic mobility (a function of length of polypeptide chain or molecular weight). SDS gel electrophoresis of samples have identical charge per unit mass due to binding of SDS results in fractionation by size.

Contents

Procedure

Tissue preparation

Samples may be taken from whole tissue or from cell culture. In most cases, solid tissues are first broken down mechanically using a blender (for larger sample volumes), using a homogenizer (smaller volumes), or by sonicator. Cells may also be broken open by one of the above mechanical methods. However, it should be noted that bacteria, virus or environmental samples can be the source of protein and thus Western blotting is not restricted to cellular studies only.

A combination of biochemical and mechanical techniques – including various types of filtration and centrifugation – can be used to separate different cell compartments and organelles.

The solution of proteins to be analyzed is mixed with SDS, an anionic detergent which denatures secondary and non–disulfide–linked tertiary structures, and applies a negative charge to each protein in proportion to its mass. Heating the samples to at least 60 degrees C shakes up the molecules, helping SDS to bind.[1] [2] [3] [4]

Full article ▸

related documents
Nucleic acid
Ytterbium
Cadmium
Fuel
Boric acid
Meitnerium
Haber process
Carbohydrate
Formic acid
Strontium
Hydronium
Nitrogen fixation
Biogas
Halogen
RNA splicing
Tellurium
Oxide
Radium
Sodium chloride
Chlorophyll
Rhodium
Scandium
Differential scanning calorimetry
Polyethylene glycol
Ultramarine
Aqua regia
Aluminium oxide
Olivine
Histone
Neptunium