The effect of including molybdate in the eluting buffer used for DEAE-cellulose chromatography on the quantitation of glucocorticoid receptor transformation.
This study analyzes the effect of including molybdate in the elution buffers used in DEAE cellulose chromatography on the fraction of glucocorticoid receptor which elutes as the transformed species. Inclusion of molybdate leads to a significant decrease in the fraction of receptor eluting as transformed; samples which appear to be nearly 50% transformed if eluted in the absence of molybdate were found to be less than 10% transformed when analyzed using a buffer which contained 5 mM molybdate. This decrease was not caused by loss of receptor or a reversion of transformation. DEAE cellulose accelerates receptor transformation. It is concluded that DEAE cellulose should not be used to quantitate transformation unless molybdate is included in all buffers.
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Department of Medicine, Louisiana State University Medical Center, New Orleans 70112.
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D000818:Animals; D002021:Buffers; D002848:Chromatography, DEAE-Cellulose / Q000379:methods; D003600:Cytosol / Q000032:analysis; D008099:Liver / Q000032:analysis; D051379:Mice; D008982:Molybdenum; D010911:Pituitary Neoplasms / Q000032:analysis; D011965:Receptors, Glucocorticoid / Q000032:analysis*
D002021:Buffers; D011965:Receptors, Glucocorticoid; C044659:molybdate; D008982:Molybdenum