Production and immunohistochemical application of antiserum against Tyr-D-Ala-Phe, a N-terminal tripeptide common to dermorphin/deltorphin family.
Tyr-D-Ala-Phe is a N-terminal sequence commonly found in a peptide family including dermorphin and deltorphin. The tripeptide was synthesized and conjugated with poly L-lysine. Nuclear magnetic resonance (NMR) indicated that approximately 38 molecules of the tripeptide were bound to each molecule of poly L-lysine. The conjugate was used to immunize rabbits, and high titer antisera were obtained. An IgG fraction was purified by protein G affinity chromatography. A specific antibody to the tripeptide was then obtained by affinity chromatography using formylcellulofine conjugated with Tyr-D-Ala-Phe. On immunospot assay, the best IgG antibody was capable of detecting 125 ng of Tyr-D-Ala-Phe but failed to react even with 2.0 microg of Tyr-L-Ala-Phe or poly L-lysine. Our immunohistochemical examination selectively localized the secretory glands of frog skin.
Tooyama
I
I
Molecular Neuroscience Research Center, Shiga University of Medical Science, Seta, 520-2192, Otsu, Japan. kinchan@sums.shiga-med.ac.jp
Abe
H
H
Renda
T G
TG
Kimura
H
H
D000409:Alanine / Q000737:chemistry*; D000818:Animals; D002846:Chromatography, Affinity; D007106:Immune Sera / Q000096:biosynthesis; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification; D007150:Immunohistochemistry; D009682:Magnetic Resonance Spectroscopy; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D018847:Opioid Peptides; D010452:Peptide Biosynthesis; D010455:Peptides / Q000737:chemistry* / Q000276:immunology; D010649:Phenylalanine / Q000737:chemistry*; D011107:Polylysine / Q000378:metabolism; D011485:Protein Binding; D011817:Rabbits; D011898:Ranidae; D012867:Skin / Q000378:metabolism; D014443:Tyrosine / Q000737:chemistry*
D007106:Immune Sera; D007074:Immunoglobulin G; D009842:Oligopeptides; D018847:Opioid Peptides; D010455:Peptides; C059567:deltorphin; D011107:Polylysine; C030058:dermorphin; D014443:Tyrosine; D010649:Phenylalanine; D000409:Alanine