Radioiodination and /sup 125/I-labeled peptide mapping of proteins on nitrocellulose membranes
A rapid procedure for generating dozens of /sup 125/I-labeled peptide maps from a protein band excised from a single lane of a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel has been developed. Proteins, which can be rapidly purified by 2 X SDS-PAGE separation, are electroblotted onto nitrocellulose paper (NCP) and located by aqueous naphthol blue-black staining. All subsequent steps of radioiodination, and enzyme or chemical cleavage, are carried out on the NCP making it possible to test a variety of cleavage reagents on the same protein sample. The resultant peptidic residues, which can be separated by thin-layer electrophoresis-thin-layer chromatography (2D TLE-TLC), SDS-PAGE, or HPLC, can be used in comparative studies or they can be recovered for further structural and immunological analyses.
- Research Organization:
- Univ. of Montana, Missoula
- OSTI ID:
- 6501031
- Journal Information:
- Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 2; ISSN ANBCA
- Country of Publication:
- United States
- Language:
- English
Similar Records
A sensitive and rapid method for identification and characterization of low abundance receptors
Detection of an azido-(/sup 14/C)-atrazine labeled protein transferred to nitrocellulose paper
Related Subjects
59 BASIC BIOLOGICAL SCIENCES
BETA DECAY RADIOISOTOPES
CELL CONSTITUENTS
CELL MEMBRANES
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
GENETIC MAPPING
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LIQUID COLUMN CHROMATOGRAPHY
MAPPING
MEMBRANES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PROTEINS
RADIOISOTOPES
SEPARATION PROCESSES
THIN-LAYER CHROMATOGRAPHY
TRACER TECHNIQUES