A rapid assay for binding estradiol to uterine receptor(s)

Erdős, T.; Best‐Belpomme, Martin; Bessada, R.

doi:10.1016/0003-2697(70)90044-8

Cited by 156 publications

(32 citation statements)

References 11 publications

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“…The concentration of the hormone receptor complexes was measured by 'differential dissociation' techniques: charcoal method [8] or hydroxylapatite method [9].…”

Section: Measurement Of Receptor Concentrationmentioning

confidence: 99%

Filtration of calf uterine cytosol on ultrogel ACA 34 prevents aggregation of the estradiol receptor

Soulignac¹,

Secco-Millet²,

Rocher³

et al. 1977

FEBS Letters

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“…The concentration of the hormone receptor complexes was measured by 'differential dissociation' techniques: charcoal method [8] or hydroxylapatite method [9].…”

Section: Measurement Of Receptor Concentrationmentioning

confidence: 99%

Filtration of calf uterine cytosol on ultrogel ACA 34 prevents aggregation of the estradiol receptor

Soulignac¹,

Secco-Millet²,

Rocher³

et al. 1977

FEBS Letters

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“…Ci/mmol; 1 Ci = 37 GBq; New England Nuclear) by using the Bolton and Hunter reagent (11 (13). Specific immunocomplexes were measured by the double-immunoprecipitation technique described above.…”

mentioning

confidence: 99%

Monoclonal antibody to chicken oviduct progesterone receptor.

Radanyi¹,

Joab²,

Renoir³

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

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Antibodies to molybdate-stabilized chicken oviduct progesterone receptor were raised in a Wistar rat and detected by interaction with homogeneous radioiodinated progesterone receptor. Spleen cells of this rat were then fused with mouse Sp2/0-Agl4 myeloma cells and the antibodies produced by the hybrid cells were detected by double immunoprecipitation using the '5I-labeled receptor. Cells of one of the positive cultures were then cloned by limiting dilution and one hybridoma cell line was studied. The monoclonal antibody produced was an IgG2b, and it reacted with the molybdate-stabilized "8-9S" form of the chicken oviduct progesterone receptor, labeled with either [H] Although the chicken oviduct progesterone receptor (PR) has been extensively studied (for review, see ref. 1), initial attempts to obtain PR antibodies were unsuccessful (2). Now, however, a homogeneous molybdate-stabilized "8-9S" form of PR has become available (3) and we have obtained anti-PR antibodies in a rabbit and a goat (4). The goat antiserum crossreacted not only with mammalian PRs (4) but, more surprisingly, with chicken glucocorticosteroid receptor (5). The latter unexpected result prompted us to obtain monoclonal antibodies (6) Exponentially growing mouse myeloma cells were fused with spleen cells by using 50% (wt/vol) polyethylene glycol 4000 and 5% dimethyl sulfoxide according to a published procedure (10) with minor modifications: 5 x 107 myeloma cells were exposed to 4 x 108 spleen cells and plated in selective medium (50 pkM hypoxanthine/10 tuM azaserine). The next day, surviving cells were counted and 0.1-ml aliquots containing 104 cells were plated in the presence of 4 X 105 rat macrophages. The medium was changed once a week. Two to 3 wk later, when the cell concentration had reached 105/ml, growing cells were tested for antibody production.The antibody-secreting hybrids were cloned by limiting dilution in the presence of 2 x 106 mouse thymocytes per ml.Wells containing a single cell cluster were assayed for anti-PR antibody production. Labeling of Chicken Oviduct Progesterone Receptor. Iodination. Highly purified (>90% pure) molybdate-stabilized PR (6 Ag) was labeled with 1 mCi of 125I (specific activity, -2,200Ci/mmol; 1 Ci = 37 GBq; New England Nuclear) by using the Bolton and Hunter reagent (11). Human Mammary Cancer Cytosol. PR-containing cytosol was prepared from MCF-7 cells (12) and total tumor, as described (4 2854The publication costs ofthis article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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“…1 M of triamcinolone acetonide was added to the culture medium 1 h before harvesting cells, and crude nuclear extracts were prepared under high ionic strength conditions (36 -38). The recombinant glucocorticoid receptor (BachGR␣) recovered in the crude nuclear extracts was analyzed for its molecular weight, and its ability to bind glucocorticoids was assessed by the hydroxyapatite method (39). The binding capacities of the crude nuclear extracts used hereafter were 3,800 pmol/ml (specific activity, 576 pmol/mg protein) or 1,710 pmol/ml (specific activity, 123 pmol/mg protein).…”

Section: Methodsmentioning

confidence: 99%

The Glucocorticoid Response Element II Is Functionally Homologous in Rat and Human Insulin-like Growth Factor-binding Protein-1 Promoters

Schweizer-Groyer¹,

Jibard²,

Neau³

et al. 1999

Journal of Biological Chemistry

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In vivo, insulin-like growth factor-binding protein-1 (IGFBP-1) modulates the IGFs' bioavailability and may contribute to their delivery to peripheral tissues. In rat and human hepatocytes, glucocorticoids stimulate IG-FBP-1 gene transcription through homologous glucocorticoid response units (GRU). Transfection experiments have shown that one of these, GRU2 (nucleotide (nt) ؊121 to ؊85 and nt ؊111 to ؊74 in human and rat promoters, respectively), was on its own able to mediate the glucocorticoid response in rat but not in human species ( . A close comparison of GRU2 sequences has pointed out a C to A transition in the underlying GREII, which creates a GATC tetranucleotide in rat species. This tetranucleotide is submitted to adenosyl methylation (dam methylation) in most Escherichia coli bacterial strains, but not in eucaryotic cells. We showed (i) that on its own, the unmethylated rat GRU2 (propagated in dam E. coli strains) was inactive, as is the case for its human counterpart (nonsignificant glucocorticoid inductions, 1.48 ؎ 0.23 and 1.7 ؎ 0.35-fold in Chinese hamster ovary cells, respectively) and (ii) that its adenosyl methylation in standard dam ؉ bacterial strains yielded a functional GRU (6.5 ؎ 1.1 and 13.1 ؎ 3.9-fold glucocorticoid inductions in Chinese hamster ovary and HepG2 cells, respectively). Transient transfection in HepG2 hepatoma cells clearly showed that the interaction of liver-enriched trans-acting factor(s) with the 5-overlapping insulin response element does not enable the unmethylated rat GRU2 or the human GRU2 to become responsive to glucocorticoids (nonsignificant 2.21 ؎ 0.48 and 1.20 ؎ 0.06-fold induction, respectively). Furthermore, we have correlated these functional data with in vitro DNA-protein interaction studies: the dam methylated rat GREII displayed a 2.8-fold higher affinity for the glucocorticoid receptor than its unmethylated counterpart.Insulin-like growth factor-binding protein-1 (IGFBP-1) 1 belongs to a family of six related proteins that bind the IGFs (IGF-I and -II) with high affinity and thus modulate their bioavailability both in the serum and in extracellular fluids, inhibit or potentiate their actions and possibly confer tissue specificity (1, 2) (also reviewed in Ref.3).In vivo, serum IGFBP-1 is thought to play an important role in the short term modulation of IGFs' bioavailability. Accordingly, its abundance is rapidly regulated and may vary by more than 10-fold in normal subjects (4). In addition, perfusion studies point out a role for circulating IGFBP-1 to deliver IGFs to peripheral tissues (5). During the perinatal period and in adults, serum IGFBP-1 is primarily synthesized in hepatocytes (6 -9). In vivo and in vitro, the hepatic production of IGFBP-1 is regulated by multiple factors (glucocorticoids, cAMP agonists, insulin, and growth hormone) (10 -12) and appears to be correlated to the abundance of its mRNA; IGFBP-1 transcripts increase after glucocorticoid treatment and in diabetic animals and decrease after insulin treatment (13-15).Regulation of IGFBP-...

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A rapid assay for binding estradiol to uterine receptor(s)

Cited by 156 publications

References 11 publications

Filtration of calf uterine cytosol on ultrogel ACA 34 prevents aggregation of the estradiol receptor

Filtration of calf uterine cytosol on ultrogel ACA 34 prevents aggregation of the estradiol receptor

Monoclonal antibody to chicken oviduct progesterone receptor.

The Glucocorticoid Response Element II Is Functionally Homologous in Rat and Human Insulin-like Growth Factor-binding Protein-1 Promoters

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