A polyclonal antibody against synthetic peptide conserved in N-Myc protein reacts with water-soluble recombinant N-Myc protein.
The importance of determining the N-Myc protein has been emphasized in neuroblastoma. We attempted to obtain a water-soluble N-Myc protein, and an antibody highly specific for the N-Myc protein. A plasmid was constructed from partial exons 2 and 3 of the N-myc gene, and expressed in Escherichia coli by isopropyl-beta,-D-thiogalactopyranoside. Newly obtained N-Myc (rN-Myc) was water-soluble with a M.W. of 38 kDa. An N-Myc-specific peptide, GVAPPRPGG RQTSGGDH, was used to raise an antibody. Specificity of the obtained antibody was confirmed and we found the rN-Myc protein reacts positively with the anti-N-Myc IgG raised here. The rN-Myc protein and the anti-N-Myc IgG obtained are expected to be used in an ELISA for N-Myc.
Yang
H W
HW
Department of Pediatric Surgery, Faculty of Medicine, University of Tokyo, Bunkyo-ku, Tokyo 113-8655, Japan.
Hemmi
H
H
Ikeda
H
H
Kato
K
K
Tsuchida
Y
Y
D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D005091:Exons / Q000235:genetics; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology / Q000302:isolation & purification; D008175:Lung Neoplasms / Q000276:immunology / Q000473:pathology; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000276:immunology; D009447:Neuroblastoma / Q000276:immunology / Q000473:pathology; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology*; D016271:Proto-Oncogene Proteins c-myc / Q000737:chemistry / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology; D012995:Solubility; D014407:Tumor Cells, Cultured; D014867:Water
D000906:Antibodies; D007074:Immunoglobulin G; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D016271:Proto-Oncogene Proteins c-myc; D011993:Recombinant Fusion Proteins; D014867:Water