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of electrophoretic techniques used nowadays involve either agarose gels or polyacryl- amide gels. 10.2.1 Agarose gels Agarose is ...

thus, for example, DNA samples separated in a gel can be cut out of the gel, returned to solution and recovered. Owing to the po ...

If this free radical is represented as R(where the dot represents an unpaired electron) and M as an acrylamide monomer molecule ...

of proteins (Section 10.3) and for the separation of DNA fragments during DNA sequence analysis (Section 10.4). Although some wo ...

buffer. When the current is switched on, all the ionic species have to migrate at the same speed otherwise there would be a brea ...

TheMrof a protein can be determined by comparing its mobility with those of a number of standard proteins of knownMrthat are run ...

relative molecular mass of the protein can be determined on the same gel run (as described above), with no more material being u ...

charges and sizes of proteins in a given protein mixture, good resolution is achieved. The enzyme of interest can be identified ...

Separation is achieved by applying a potential difference across a gel that contains a pH gradient. The pH gradient is formed by ...

charge on the protein will decrease correspondingly until eventually the protein arrives at a point where the pH is equal to its ...

10.3.5 Two-dimensional polyacrylamide gel electrophoresis This technique combines the technique of IEF (first dimension), which ...

2-D gels (20 cm20 cm), although the minigel system can be used to provide useful separation in some cases. For large-format gel ...

soaked in buffer and substrate. After an appropriate incubation period, the strips are peeled apart and the paper zymogram treat ...

10.3.7 Detection, estimation and recovery of proteins in gels The most commonly used general protein stain for detecting protein ...

approach, the major bands on the gel can be identified with CBB and then minor bands, not detected with CBB, resolved using the ...

(electroelution). A number of different designs of electroelution cells are commercially available, but perhaps the easiest meth ...

protein. The blot is first incubated in a protein solution, for example 10% (w/v) bovine serum albumin, or 5% (w/v) non-fat drie ...

horseradish peroxidase is to use the method of enhanced chemiluminescence (ECL). In the presence of hydrogen peroxide and the ch ...

Quantum dots: These are engineered semiconductor nanoparticles, with diameters of the order of 210 nm, which fluoresce when ex ...

and for the more flexible single-stranded DNA. It will be obvious from the above that gel concentrations must be chosen to suit ...