manufacturer :       ancos

model :                   Semi-Dry B

annotation :           Dokumente engl .

For modern biologists the identification of proteins in complex mixtures is the basis for their subsequent characterization and isolation. By means of the high resolution SDS- polyacryl­amide gel electrophoresis, especially in its 2 dimensional version, it is possible to resolve virtually any mixture of proteins. However, further analysis of the separated proteins is limited due to their relative inaccessibility within the gel matrix. lt was therefore a major contribution to the techniques available, when the concept of transferring the fractionated proteins out of the gel matrix, to an immobilized state on a membrane was developed. By this technique, termed blotting, the proteins simultaneously maintain their resolution and become accessible to a wide variety of detection means and further analyses.

The main advantages of the blotting principle are due to:

1. Accessibility

The antigens are concentrated at the surface of the membrane texture.

2. Immobilization

Since diffusion is avoided the original resolution is preserved.

3. Time

Semi-dry electroblotting can be performed within one hour.

4. Economics

Small amounts of proteins are required (10 - 100 ng ) as a high recovery on the blotting membrane is obtained.

5. Versality

Depending on the technique employed for the protein separation denatured or native proteins can be analysed . Thus, in contrast to the denaturing appearing when SDS-containing gels are used, native protein can be separated for example by conventional zone electrophoresis isoelectrofocusing in agarose and polyacrylamide gel or crossed immunoelectrophoresis . Also material from thin layer chromatography can be blotted.

6. Flexibliity

The blots might be stored and reused by probing with different ligands . The principle is applicable to all types of ligands (e.g. antigens/antibodies, receptors/- hormones, glycoproteins/lectins ).