Characterization of polyclonal antibodies against the N-terminal domain of the human androgen receptor.
Antibodies against the N-terminal domain of the human androgen receptor (hAR) were prepared by two different approaches. Firstly, rabbits were immunized with a beta-galactosidase-hAR (amino acids (aa) 174-353) fusion protein. Secondly, two synthetic peptides corresponding to potentially antigenic sites located within this fragment (aa 201-222 and 301-320) were used as immunogens. The obtained antisera contained high titer anti-hAR antibodies as was established with several independent methods (e.g. sucrose gradient centrifugation, immunoprecipitation, Western blotting). The two anti-peptide antisera specifically stained nuclei of glandular epithelial cells in frozen sections of human prostate tissue. Progesterone, estradiol and glucocorticoid receptors were not immunoprecipitated with these antisera. The specific hAR antibodies provide new tools for the characterization of this steroid receptor as well as for diagnostic purposes in pathology of the human prostate and androgen resistance.
van Laar
J H
JH
Department of Biochemistry II, Erasmus University Rotterdam, The Netherlands.
Voorhorst-Ogink
M M
MM
Zegers
N D
ND
Boersma
W J
WJ
Claassen
E
E
van der Korput
J A
JA
Ruizeveld de Winter
J A
JA
van der Kwast
T H
TH
Mulder
E
E
Trapman
J
J
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D002467:Cell Nucleus / Q000032:analysis; D000939:Epitopes / Q000276:immunology; D006801:Humans; D008297:Male; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology; D011467:Prostate / Q000032:analysis / Q000648:ultrastructure; D011817:Rabbits; D011944:Receptors, Androgen / Q000276:immunology*; D011987:Receptors, Steroid / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000276:immunology
D000906:Antibodies; D000939:Epitopes; D010446:Peptide Fragments; D011944:Receptors, Androgen; D011987:Receptors, Steroid; D011993:Recombinant Fusion Proteins