geelelektrofreesil lahutatud sarnaste makromolekulide identifitseerimiseks kasutatav tehnika, mis seisneb komponentide ülekandes geelist spetsiaalsele membraanile ja nende järgnevas detekteerimises
technique used to detect the presence of a polypeptide which is antigenic to the specific antibody used in the assay. A protein mixture is denaturated and separated on the basis of size by SDS gel electrophoresis, then blotted on to a sheet of nitrocellulose, preserving the pattern due to size. The proteins in the pattern are reacted with the specific antibody, then with an AP- or HRP-conjugated second antibody against the first which after the development, produces visible staining at the site of binding.
The following information on the library from which the insert originated, should be known:
(i) the source and method for obtaining the nucleic acid of interest (cDNA, chromosomal, mitochondrial, etc.);
(ii) the vector in which the library was constructed (e.g. lamda GT 11, pBR 322, etc.) and the site in which the DNA was inserted;
(iii) the method used for identification (colony, hybridization, immuno-blot, etc.);
(iv) the strain used for library construction.
