By M. Dean Savage (auth.), Dr. Thomas Meier, Dr. Falk Fahrenholz (eds.)
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Extra resources for A Laboratory Guide to Biotin-Labeling in Biomolecule Analysis
1994) Anal. Biochem. 218, 330-337. D. and Shugar, D. (1964) Photochemistry of Proteins and Nucleic Acids. Pergamon Press, Oxford. Purification of the Receptor for Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) using Biotinylated Ligands Tetsuya Ohtaki, Chieko Kitada and Haruo Onda Summary Th pituitary adenylate cyclase-activating polypeptide (PACAP) including PACAP27 and PACAP38, timulates adenylate cycla e via specific PACAP receptors that are coupled to G protein. The pre ent tudy describes the application of avidin-biotin technology to the purification of the PACAP receptor from bovine brain membrane.
Biochem. 138, 141-143. P. and Hancock, R. (1981) Anal. Biochem. 118, 197-203. , Hughes, J. R. (1989) Eur. J. Pharmacol. 172, 339-346. B. (1970) J. Am. Chern. Soc. 92,6333 . H. K. (1975) J. Am. Chern. Soc. 97, 1575-1579. , Albericio, F. and Barany, G. (1990) Int. J. Peptide Protein Res. 35,527-538. , Stueber, W. and Bayer, E. (1982) Physiol. Chern. Hoppe-Seyler 363, 701-708. Thiele, e. and Fahrenholz, F. (1994) Anal. Biochem. 218, 330-337. D. and Shugar, D. (1964) Photochemistry of Proteins and Nucleic Acids.
Purification was performed in 3 sequential steps. First, antiserum was incubated with [3H]BANA-CCK -8s. The amount of nonspecific binding was determined by a parallel experiment where 10 flM pentagastrin was added at this stage. Alternatively, preimmune serum was used to distinguish specific from nonspecific binding. Then streptavidin-agarose beads were added, and the suspension was equilibrated for 3 hours at 4 DC to allow complete binding of the excess of biotinylated ligand and of the ligand-antibody complex to the matrix.
A Laboratory Guide to Biotin-Labeling in Biomolecule Analysis by M. Dean Savage (auth.), Dr. Thomas Meier, Dr. Falk Fahrenholz (eds.)