Immunohistochemical detection of the human cytochrome P4507B1: production of a monoclonal antibody after cDNA immunization.
The cytochrome P4507B1 (P4507B1) is responsible for the 7alpha-hydroxylation of dehydroepiandrosterone (DHEA) and other 3beta-hydroxysteroids in the brain and other organs. The cDNA of human P4507B1 was used for DNA immunization of mice. The best responding mouse led to the production of monoclonal antibodies (mAbs). The clone D16-37 produced an IgM specific for P4507B1 with no cross-reaction with other human P450s. This antibody permitted the immunohistochemical detection of P4507B1 in slices of human hippocampus. P4507B1 was expressed in neurons only. This new tool will be used for the extensive examination of the P4507B1 presence and determination of its levels in slices of human normal and diseased brain and in other human tissues.
Trap
Catherine
C
Laboratoire de Biotechnologie, EA-3199, Conservatoire National des Arts et Métiers, 2 rue Conté, 75003 Paris, France.
Nato
Farida
F
Chalbot
Sonia
S
Kim
Sae-Bom
SB
Lafaye
Pierre
P
Morfin
Robert
R
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000096:biosynthesis / Q000378:metabolism; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D002384:Catalysis; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis* / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000072474:Cytochrome P450 Family 7; D018076:DNA, Complementary / Q000008:administration & dosage* / Q000276:immunology; D003687:Dehydroepiandrosterone / Q000037:antagonists & inhibitors / Q000378:metabolism; D006624:Hippocampus / Q000201:enzymology / Q000276:immunology; D006801:Humans; D007075:Immunoglobulin M / Q000378:metabolism; D007150:Immunohistochemistry; D007273:Injections, Intramuscular; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D013250:Steroid Hydroxylases / Q000032:analysis* / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000276:immunology
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D007075:Immunoglobulin M; D019444:Vaccines, DNA; D003687:Dehydroepiandrosterone; D003577:Cytochrome P-450 Enzyme System; D013250:Steroid Hydroxylases; C115867:oxysterol 7-alpha-hydroxylase; D000072474:Cytochrome P450 Family 7; C499571:CYP7B1 protein, human