公开(公告)号：JPS63206658A

公开(公告)日：1988-08-25

申请号：JP3995387

申请日：1987-02-23

Applicant: AGENCY IND SCIENCE TECHN

Inventor： YOKOYAMA NAOKI ,  NAKAO YUKIMICHI ,  OHASHI SHINICHI ,  KAERIYAMA KYOJI ,  TSUDA KEISHIRO ,  SUDA MASAO

IPC: G01N33/68

Abstract: PURPOSE:To safely and economically detect the protein immobilized on a membrane with high sensitivity, by adsorbing palladium colloid by protein and applying the electroless plating of a base metal to said protein to sensitize the same. CONSTITUTION:In detecting the protein immobilized on a membrane, this membrane is treated with palladium colloid stabilized with a nonionic surfactant. As the membrane used in the immobilization of protein to be detected, any one commonly used in a gold colloid method is used. As a method for immobilizing protein to be inspected on the membrane, for example, an electroblotting method is used. Subsequently, the electroless plating of a base metal is further applied to the membrane thus treated to sensitize the protein. This electroless plating is a catalytic chemical plating utilizing the catalytic activity of palladium and a metal is selectively precipitated on a metal surface having catalytic action. By applying immersion treatment to the treated membrane in an electroless plating liquid, the base metal is precipitated on palladium colloidal particles from said plating liquid and, therefore, protein can be detected with high sensitivity.

公开(公告)号：JPS63111465A

公开(公告)日：1988-05-16

申请号：JP25719086

申请日：1986-10-29

Applicant: AGENCY IND SCIENCE TECHN

Inventor： KOKUBU TOMOKUNI ,  IWAKURA MASAHIRO ,  OHASHI SHINICHI ,  TSUDA KEISHIRO

IPC: C12N15/02 ,  C12N15/00 ,  C12P21/00 ,  C12Q1/00 ,  G01N33/53 ,  G01N33/535 ,  G01N33/577

Abstract: PURPOSE:To improve measurement sensitivity and reproducibility by utilizing the fused protein genetically bonded with labeling enzyme and antigen by a gene manipulation as a labeling antibody. CONSTITUTION:The coliform bacilli contg. the plasmid having the gene coding the dihydro folic acid reductase-leucine enkephalline fused protein (hereafter expressed as DHFR-L-Enk) are prepd. by the gene manipulation and are subjected to fractionating, refining and purifying after liquid culture to obtain the DHFR-L-Enk exhibiting a uniform band in SDS electrophoresis. Said liquid is added together with an antirabbit leucine enkephalline (hereafter expressed as L-Enk) antibody to cause reaction, by which a soln. for measuring enzyme activity is obtd. A DHFR substrate soln. is then added thereto to initiate reaction. A change in the absorption at 340nm wavelength is measured by a spectrophotometer, by which L-Enk is measured.