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US 4,784,960 AGranted
Process for preparation of control for use in progesterone receptor
AssigneePeter S. Brune
InventorRichard J. Baranczuk
Life of this patent
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Abstract
A process is disclosed for producing steroid hormone receptor samples to be utilized as controls during assays of various human tissue for steroid hormones, especially estrogen and progesterone. The process comprises collecting tissue known to include such receptors, adding a buffer solution to the tissue, homogenizing the tissue and buffer solution, centrifuging the homogenized mixture, and thereafter collecting the supernatant. The supernatant which contains the desired receptors is subdivided into suitable control sample size and preferably lyophilized to a flake. A process is described for maximizing the number of receptors found in the samples.
Claims21
A process for production of a control sample for use in progestin receptor assays;
- said process comprising the steps of:
- (a) inducing superovulation in a female mammal in conjunction with mating of the mammal so as to produce pregnancy within the mammal;
- (b) thereafter collecting a mammalian tissue from said mammal containing progestin receptors;
- (c) adding a buffer solution to said tissue;
- (d) homogenizing said tissue and said buffer together;
- (e) centrifuging said homogenized tissue and buffer;
- and
- (f) collecting the supernatant from said centrifuged tissue and buffer as said control sample.
5 more elements
A process for the production of a control sample for use in progesterone receptor assays;
- said process comprising the steps of:
- (a) inducing superovulation in rabbits;
- (b) in conjunction with such superovulation inducing pregnancy in the rabbits;
- (c) collecting uteri tissue from approximately 6 day pregnant rabbits following steps (a) and (b);
- (d) freezing said tissue;
- (e) shreading said tissue into relatively small strips;
- (f) adding a buffer solution including sodium molybdate and having a pH of approximately 7.5 to said tissue;
- (g) homogenizing said buffer and tissue;
- (h) centrifuging said homogenized buffer and tissue;
- (i) removing the supernatant from said centrifuged buffer and tissue;
- (j) freezing said supernatant;
- and
- (k) subjecting said frozen supernatant to a vacuum so as to freeze dry said supernatant into dry flakes.
10 more elements
A receptor process for the production of a control sample for use in progesterone receptor assays;
- said process comprising the steps of:
- (a) inducing superovulation in rabbits in conjunction with inducing pregnancy in such rabbits;
- (b) collecting utrei tissue from such rabbits after the rabbits have been pregnant for a period of approximately 6 days;
- (c) freezing said tissue to less than approximately -70 degrees C.;
- (d) shreading said tissue into relatively small strips;
- (e) adding a buffer solution including sodium molybdate and having a pH of approximately 7.5 to said tissue;
- (f) homogenizing said buffer and tissue in a blender for periods of less than approximately ten seconds while allowing the tissue and buffer to be cooled between periods of homogenization until substantial homogenization occurs;
- (g) centrifuging said homogenized buffer and tissue;
- (h) removing the supernatant from said centrifuged buffer and tissue;
- (i) freezing said suprenatant to less than about -70 degrees C.;
- (j) placing said supernatant in a plurality of approximately equal aliquots;
- (k) subjecting said frozen supernatant to a vacuum so as to freeze dry said supernatant into dry flakes;
- and
- (1) placing said dry flakes after freeze drying into a vacuum retaining container for storage until use as a control sample.
11 more elements
A process for production of a control sample for use in progestin receptor assays;
- said process comprising the steps of:
- (a) collecting a mammalian tissue comprising tissue from the group consisting essentially of normal breast, ovaries, fallopian tubes, uterus, placenta, corpus lutem tissues and combinations thereof;
- said mammalian tissue containing progestin receptors from a female animal selected on approximately a uniform given day of a female cycle associated with such animal, such that the progestin receptor level in said tissue is relatively constant for each animal species;
- (b) adding a buffer solution to said tissue such that the pH of said tissue after adding said buffer is in the range of approximately 6 to 8;
- (c) adding a progestin receptor stabilizing agent to said tissue;
- (d) homogenizing said tissue, buffer and said agent together;
- (e) centrifuging said homogenized tissue, buffer and agent;
- (f) collecting the supernatant from said centrifuged tissue and buffer as said control sample;
- and
- (g) freeze drying said control sample.
7 more elements
A process for the production of a control sample for use in progesterone receptor assays;
- said process comprising the steps of:
- (a) collecting uteri tissue from approximately 6 day pregnant rabbits;
- (b) freezing said tissue;
- (c) shreading said tissue into relatively small strips;
- (d) adding a buffer solution having a pH of approximately 7.5 to said tissue;
- (e) adding a progesterone receptor stabiliziing agent to said tissue;
- (f) homogenizing said buffer, agent and tissue;
- (g) thereafter centrifuging said buffer, agent and tissue for approximately one half hour at a relatively high RPM;
- (h) removing the supernatant from said centrifuged buffer, agent and tissue;
- (i) freezing said supernatant;
- and
- (j) subjecting said frozen supernatant to a vacuum so as to freeze dry said supernatant into dry flakes.
9 more elements
A process for the production of a control sample for use in progesterone receptor assays;
- said process comprising the steps of:
- (a) collecting uteri tissue from approximately 6 day pregnant rabbits;
- (b) freezing said tissue to less than approximately -70 degrees C.;
- (c) shreading said tissue into relatively small strips;
- (d) adding a buffer solution including sodium molybdate and having a pH of approximately 7.5 to said tissue;
- (e) homogenizing said buffer and tissue in a blender for periods of less than ten seconds while allowing the tissue and buffer to be cooled between periods of homogenization until substantial homogenization occurs;
- (f) centrifuging said homogenized buffer and tissue for approximately one half hour at approximately 40,000 RPM;
- (g) removing the supernatant from said centrifuged buffer and tissue;
- (h) freezing said supernatant to less than about -70 degrees C.;
- (i) placing said supernatant in a plurality of approximately equal aliquots;
- (j) subjecting said frozen supernatant to a vacuum so as to freeze dry said supernatant into dry flakes;
- and
- (k) placing said dry flakes after freeze drying into a vacuum retaining container for storage until use as a control sample.
10 more elements
A process for the production of a control sample for use in steroid hormone receptor assays;
- said process comprising the steps of:
- (a) collecting normal mammalian tissue having a concentration of said steroid hormone therein;
- (b) freezing said tissue;
- (c) shreading said tissue into relatively small strips;
- (d) adding a buffer solution having a pH of approximately 7.5 and sodium molybdate to said tissue;
- (e) homogenizing said buffer and tissue;
- (f) thereafter centrifuging said buffer and tissue for sufficient time at a relatively high RPM to separate a portion of said buffer and tissue into a supernatant;
- (g) removing said supernatant from said centrifuged buffer and tissue;
- (h) freezing said supernatant;
- and
- (i) subjecting said frozen supernatant to a vacuum so as to freeze dry said supernatant into dry flakes.
8 more elements
Dependencies are inferred from the claim text.