Multicolour immuno-staining of fibrinogen polypeptide chains for identification of their derivatives in electrophoregrams.
A novel electrophoretic procedure enabling multiple, direct immunoprobing of electrophoregrams without depending on Western blotting is described, and applied to the identification of the derivatives formed in the early stages of clot stabilization. Multicolour immunostainings for positive identification of cross-linked chains in partially stabilized fibrin clots indicated that the early products of alpha-chain cross-linking by factor XIII are largely hybrids of co-cross-linking of alpha- and gamma-chains rather than alpha-chain polymers suggested from previous studies employing non-specific staining of electrophoregrams. Furthermore, plasma-fibrinogen dimers were found to contain cross-linked alpha-chains with an electrophoretic mobility very near that of gamma-gamma-dyads. A similar product is produced by tissue-transglutaminase, but not by factor XIII.
Shainoff
J R
JR
Research Institute, Cleveland Clinic Foundation, Ohio 44195.
Urbanic
D A
DA
D015100:3,3'-Diaminobenzidine; D002021:Buffers; D002227:Carbazoles; D004587:Electrophoresis, Agar Gel / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005340:Fibrinogen / Q000032:analysis*; D006038:Glyoxylates; D006801:Humans; D007122:Immunoelectrophoresis / Q000379:methods*; D007124:Immunoenzyme Techniques; D009284:Naphthols; D010455:Peptides / Q000032:analysis; D012685:Sepharose; D013194:Staining and Labeling
D002021:Buffers; D002227:Carbazoles; D006038:Glyoxylates; D009284:Naphthols; D010455:Peptides; C070399:glyoxyl agarose; D015100:3,3'-Diaminobenzidine; C025917:4-chloro-1-naphthol; C020702:3-amino-9-ethylcarbazole; D005340:Fibrinogen; D012685:Sepharose