Method for determining follicle stimulating hormone
First Claim
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1. In a method for the determination of follicle stimulating hormone in a sample, comprising the steps of:
- (a) contacting said sample with a purified follicle stimulating hormone receptor preparation capable of selectively binding said hormone;
(b) providing a signalling entity for signalling whether said binding has taken place; and
(c) observing said signalling entity to determine the presence of said hormone in said sample,the improvement comprising said signalling entity being a substantially pure follicle stimulating hormone labeled with a detectable label, in which labeled hormone the ratio of specific binding to non-specific binding is at least about 5 when measured using a partially purified calf testes plasma membrane preparation containing from about 5 to about 25 fmol of receptors per milligram of membrane wet weight, and said purified follicle stimulating hormone receptor preparation having at least about 600 fmol/mg protein of high affinity receptors for follicle stimulating hormone.
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Abstract
The present disclosure relates to an improved method based on hormone-receptor binding for the determination of follicle stimulating hormone and to improved reagents useful for the determination of follicle stimulating hormone in a hormone-receptor binding assay.
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Citations
9 Claims
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1. In a method for the determination of follicle stimulating hormone in a sample, comprising the steps of:
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(a) contacting said sample with a purified follicle stimulating hormone receptor preparation capable of selectively binding said hormone; (b) providing a signalling entity for signalling whether said binding has taken place; and (c) observing said signalling entity to determine the presence of said hormone in said sample, the improvement comprising said signalling entity being a substantially pure follicle stimulating hormone labeled with a detectable label, in which labeled hormone the ratio of specific binding to non-specific binding is at least about 5 when measured using a partially purified calf testes plasma membrane preparation containing from about 5 to about 25 fmol of receptors per milligram of membrane wet weight, and said purified follicle stimulating hormone receptor preparation having at least about 600 fmol/mg protein of high affinity receptors for follicle stimulating hormone. - View Dependent Claims (3, 4)
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2. In a method for the determination of follicle stimulating hormone in a sample, comprising the steps of:
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(a) contacting said sample with a purified follicle stimulating hormone receptor preparation capable of selectively binding said hormone; (b) providing a signalling entity for signalling whether said binding has taken place; and (c) observing said signalling entity to determine the presence of said hormone in said sample, the improvement comprising said signalling entity being a substantially pure follicle stimulating hormone labeled with a detectable label, prepared from an unpurified labeled follicle stimulating hormone and characterized as follows; (i) the substantially pure labeled follicle stimulating hormone co-migrates as a single band with unlabeled follicle stimulating hormone when electrophoresed on 7.5% polyacrylamide gel with an acrylamide to BIS ratio of about 19;
1 in an electrophoresis buffer containing 9.3 g of tris(hydroxymethyl)tetraacetic acid (disodium salt), 0.7 g of boric acid and 5 mg of merthiolate per liter of water at a pH of from 8.9 to 10; and(ii) when the unpurified, labeled follicle stimulating hormone is chromatographed on a G-25 column and a portion of the labeled follicle stimulating hormone of any fraction from the column having the same amount of detectable label as used in part (i) is electrophoresed as described in part (i), the amount of detectable label found in the single band of part (i) is at least about three times the amount of detectable label in the co-migrating band of material from the fraction of the column; and said purified follicle stimulating hormone receptor preparation having at least about 600 fmol-/mg protein of high affinity receptors for follicle stimulating hormone.
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- 5. The method of claim wherein said radioactive label is 125 I or 131 I.
Specification