A method for selectively staining water soluble protein using reactive dye

摘要

Staining water-soluble protein by reacting reactive dye e.g., triazine series reactive dye, with water-soluble protein for carrying out staining reaction in acidic or alkaline environment, preferably in acidic environment if protein is antibody, dye reacts with hydroxy group from amino acid side chain of protein, hydroxy group from constant region or heavy chain of antibody, hydroxy group from oligosaccharide coupled to antibody. Staining (M1) a water-soluble protein involves reacting a reactive dye with a water-soluble protein for carrying out a staining reaction, where the reactive dye is chosen from a triazine series reactive dye and a vinyl sulfone series reactive dye or mixed triazine and vinyl sulfone series reactive dye used in a textile industry, and the staining reaction is carried out in an acidic or alkaline environment, where the staining is carried out in the acidic environment if the water-soluble protein is glycoprotein, preferably antibody, so that the dye would react with a hydroxy group from an amino acid side chain of the protein, a hydroxy group from the constant region of the antibody, a hydroxy group from heavy chain of the antibody and a hydroxy group from an oligosaccharide conjugated to the antibody. An independent claim is also included for staining (M2) selectively a glycoprotein particularly an antibody, involves reacting a reactive dye with a natural or synthetic water-soluble polymer for carrying out the conjugation of the dye-loaded carrier to the oligosaccharide moiety of the antibody molecule selectively after periodate activation treatment, the staining reaction being carried out in an alkaline environment, the conjugation reaction being carried out in an appropriate covalent bond forming environment, where the staining reaction is carried out in the alkaline medium if the water-soluble polymer is a natural or synthetic water-soluble polymer, so that a dye molecule would react with a hydroxy group or an amino group from a natural or synthetic water-soluble polymer then the stained polymer can be conjugated to the activated oligosaccharide moiety of antibody molecules in an appropriate covalent bond-forming reaction, where the staining reaction is carried out to the natural or synthetic water-soluble polymeric carrier before it is conjugated to antibody molecule, so that a dye molecule would react with a electron riched functional group in the natural or synthetic water-soluble polymer, in any feasible number to enhance staining intensity such as immunoassay sensitivities, under manipulatable conditions without interfering the antigen-antibody reaction.