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Immunocytochemical application of monoclonal antibodies to rat liver glucocorticoid receptor
Author(s) -
Teasdale Jane,
Lewis Fraser A.,
Barrett Ivor D.,
Abbott Alison C.,
Wharton John,
Bird Colin C.
Publication year - 1986
Publication title -
the journal of pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.964
H-Index - 184
eISSN - 1096-9896
pISSN - 0022-3417
DOI - 10.1002/path.1711500402
Subject(s) - glucocorticoid receptor , monoclonal antibody , immunoperoxidase , receptor , biology , glucocorticoid , immunostaining , immunohistochemistry , microbiology and biotechnology , staining , antibody , endocrinology , medicine , immunology , biochemistry , genetics
Seven per cent (10/145) of hybridomas raised against partially purified activated glucocorticoid receptor from rat liver produced monoclonal antibody to receptor. Six IgM secreting clones selected for further investigation bound equally well to activated and non‐activated receptor from fresh rat liver, but significantly less well (11–25 per cent) to receptor from frozen rat liver. No interaction was found with oestrogen receptor from rat uterus but extensive cross reaction occurred with progesterone receptor. Although none of the antibodies bound to glucocorticoid receptor from human or porcine liver or lymphoid cells, several cross‐reacted with mouse liver glucocorticoid receptor. Immunoelectroblotting of proteins from fresh and frozen rat liver cytosol showed the antibodies bound to 90 000 and 40 000 MW forms of receptor respectively. Immunostaining of both frozen and paraffin embedded sections of rat tissue showed that receptor is preserved during fixation and processing of tissues. Using both indirect immunoperoxidase and immunogold silver staining methods, the pattern of receptor staining observed correlates with the known glucocorticoid responsiveness of the tissues studied.