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Mol Cell Endocrinol. 2014 Mar 5;383(1-2):111-7. doi: 10.1016/j.mce.2013.11.022. Epub 2013 Dec 8.

Development of monoclonal antibodies against human CYP11B1 and CYP11B2.

Author information

1
Endocrine Section, G.V. (Sonny) Montgomery VA Medical Center, USA; Endocrinology, University of Mississippi Medical Center, USA. Electronic address: cgomez-sanchez@umc.edu.
2
Endocrinology, University of Mississippi Medical Center, USA.
3
Department of Obstetrics and Gynecology, University of Alabama, Birmingham, AL, USA.
4
Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI, USA.
5
Tohoku University, Department of Pathology, Tohoku University, Sendai, Japan.
6
Endocrine Section, G.V. (Sonny) Montgomery VA Medical Center, USA; Endocrinology, University of Mississippi Medical Center, USA; Department of Pharmacology and Toxicology, University of Mississippi Medical Center, Jackson, MS, USA.

Abstract

1. The final enzymes in the biosynthesis of aldosterone and cortisol are by the cytochrome P450 CYP11B2 and CYP11B1, respectively. The enzymes are 93% homologous at the amino acid level and specific antibodies have been difficult to generate. 2. Mice and rats were immunized with multiple peptides conjugated to various immunogenic proteins and monoclonal antibodies were generated. The only peptide sequences that generated specific antibodies were amino acids 41-52 for the CYP11B2 and amino acids 80-90 for the CYP11B1 enzyme. 3. The mouse monoclonal CYP11B2-41 was specific and sensitive for use in western blots and produced specific staining of the zona glomerulosa of normal adrenal glands. The rat monoclonal CYP11B1-80 also detected a single band by western blot and detected only the zona fasciculata. Triple immunofluorescence of the adrenal demonstrated that the CYP11B1 and the CYP11B2 did not co-localize, while as expected the CYP11B1 co-localized with the 17α-hydroxylase.

KEYWORDS:

17α-Hydroxylase; Adrenal cortex; CYP11B1; CYP11B2; Immunofluorescence; Monoclonal antibodies

PMID:
24325867
PMCID:
PMC3939805
DOI:
10.1016/j.mce.2013.11.022
[Indexed for MEDLINE]
Free PMC Article
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