Development of a monoclonal antibody against recombinant neuroendocrine 7B2 protein.
Mouse monoclonal antibody MON-100 was raised against the neuroendocrine protein 7B2 using bacterially produced hybrid proteins. In Western blot analysis, MON-100 reacted with 3 different 7B2 hybrid proteins and not with the respective carrier proteins. Furthermore, MON-100 was reactive with recombinant 7B2 cleaved from a hybrid protein. In an immunohistochemical study, MON-100 exhibited strong reactivity with the intermediate lobe of the Xenopus pituitary gland, a tissue previously shown to contain 7B2 mRNA. Using MON-100, immunoprecipitation analysis of newly synthesized proteins produced by in vitro incubated Xenopus neurointermediate lobes revealed the biosynthesis of a single protein of Mr 24 kDa, the expected size of the 7B2 protein. It appears, therefore, that the anti-7B2 monoclonal antibody MON-100 can be successfully used for Western blot analysis and immunohistochemical analysis as well as for immunoprecipitation experiments.
van Duijnhoven
H L
HL
Dept of Biochemistry, University of Nijmegen, The Netherlands.
Ayoubi
T A
TA
Timmer
E D
ED
Braks
A A
AA
Roebroek
A J
AJ
Martens
G J
GJ
van de Ven
W J
WJ
D000818:Animals; D000911:Antibodies, Monoclonal; D001921:Brain / Q000166:cytology*; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D009419:Nerve Tissue Proteins; D051859:Neuroendocrine Secretory Protein 7B2; D010902:Pituitary Gland / Q000166:cytology*; D010907:Pituitary Hormones / Q000276:immunology; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D014981:Xenopus
D000911:Antibodies, Monoclonal; D009419:Nerve Tissue Proteins; D051859:Neuroendocrine Secretory Protein 7B2; D010907:Pituitary Hormones; D011994:Recombinant Proteins