ISSN:
1573-3904
Keywords:
G-protein receptor
;
Immunohistochemistry
;
Gastrointestinal hormones
Source:
Springer Online Journal Archives 1860-2000
Topics:
Chemistry and Pharmacology
Notes:
Summary Seven synthetic peptides, between 7–22 residues long, corresponding to six different parts of the gastrin/CCKB receptor molecule which are conserved among the species, were used for raising antibodies. The peptides were coupled to keyhole limpet hemocyanine and injected into rabbits. ELISA analysis demonstrated that all peptides produced an immune response after three to six injections given at biweekly intervals. The titer ranged from 1:104 to 1:105. All antibodies recognized a 78 kDa protein on immunoblots of NIH 3T3 cells stably transfected with human gastrin/CCKB receptor cDNA, as well as human and guinea pig stomach mucosal extracts. Preincubation of the sera with the corresponding peptides abolished the staining. Indirect immunofluorescence staining revealed that four antibodies out of the seven tested recognized the receptor in fixed COS-7 cells transiently transfected with human gastrin/CCKB receptor cDNA. The reactive antibodies were raised against the peptides corresponding to receptor residues 40–58, 153–160, 288–294 and 356–372. Immunohistochemical staining of guinea pig stomach using these antisera resulted in intense staining of parietal cells in the fundus and cardia regions.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00127268
