A controlled pore glass bead assay for the measurement of cytoplasmic and nuclear glucocorticoid receptors.
An assay for the quantitation of cytoplasmic and nuclear glucocorticoid receptors in lymphoid tissue has been developed using controlled pore glass (CPG) beads. Soluble receptor--3H-steroid complex (cytosol or nuclear extract) is adsorbed quantitatively within the crevasses of porous glass beads. Excess labeled steroid as well as most non-specifically bound steroid is easily washed away, leaving the hormone-receptor complex retained by the beads. Bound 3H-steroid is eluted with ethanol and measured for radioactivity. This procedure which is simple, rapid, and highly reproducible is carried out using frozen samples (stable for many months) containing as few as 1 X 10(7) cells. A comparison of the CPG assay to dextran coated charcoal and a whole cell assay demonstrates that CPG and dextran coated charcoal give equivalent measurements of cytosolic receptor concentration, while the CPG and whole cell assays provide equivalent values for total receptor content.
Gormley
G J
GJ
Hospelhorn
V D
VD
Khan
M K
MK
Jensen
E V
EV
D000327:Adsorption; D000818:Animals; D002467:Cell Nucleus / Q000032:analysis*; D002606:Charcoal; D003345:Corticosterone / Q000378:metabolism; D003593:Cytoplasm / Q000032:analysis*; D003911:Dextrans; D005260:Female; D005898:Glass; D006801:Humans; D008214:Lymphocytes / Q000032:analysis; D008221:Lymphoid Tissue / Q000032:analysis*; D008863:Microspheres; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000032:analysis* / Q000378:metabolism; D011987:Receptors, Steroid / Q000032:analysis*; D013950:Thymus Gland / Q000032:analysis; D014222:Triamcinolone Acetonide / Q000378:metabolism
D003911:Dextrans; D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D002606:Charcoal; D014222:Triamcinolone Acetonide; D003345:Corticosterone