Environmental Engineering Reference
In-Depth Information
Fig. 15.1 Several decades of gel electrophoresis research developments and milestones as year
scale.
GE
gel electrophoresis,
IEF
isoelectric focusing,
SDS
sodium dodecyl sulphate
15.1.2 One-Dimensional Gel Electrophoresis (1-DE)
1-DE application also known as sub-marine agarose gel electrophoresis can be used
to perform separation based on one property, i.e. charge property. In general, 1D
SDS PAGE is used to analyse less complex samples and to get idea of most
abundant proteins. This technique provides feasibility to compare different samples
on a single gel. Use of silver staining provides further refinement. This technique
can be combined easily with other protein identification methods. The 1-DE
analysis is commonly employed as a fast and easy tool to identify less complex
samples. Feasibility of multiple sample analysis (up to 10) on a single gel of almost
any size is possible. Analysis of sample before and after denaturation provides info
on carbohydrate while radiolabelling can provide information on sulphate, phos-
phate and carbohydrate content. Identified protein can be recovered from gel by
electroelution, electroblotting on polybrene coated derivative glass fibre sheet or
polyvinylidene difluoride membrane filters method by applying further experimen-
tal steps [
1
]. Multiple casting of gels at one go can provide robust result by avoiding
gel-to-gel variation. Mini gel slabs are becoming increasingly popular for isolation
of peptide sequencing by taking unique advantage of speed and high resolution. The
small gels are easily adapted to single concentration gradient and two-dimensional
SDS-PAGE procedures [
2
].
15.1.3 Two-Dimensional Gel Electrophoresis 2-DE
It is abbreviated as 2-DE or 2-D electrophoresis and is a form of gel electrophoresis
commonly used to analyse proteins. This technique separates soluble proteins with
