US4684609A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Method and substance for the enhanced labelling of cellular material
Assignee: Vector Laboratories Inc
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Filing: 1981-06-15
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Granted: 1987-08-04
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Expires: 2001-06-15
Abstract
Abstract An enhanced labelling complex for localizing markers on a prepared tissue section is disclosed. The complex includes both avidin components and biotinylated macromolecular components at a ratio preselected to provide a complex which is sufficiently large to include a large number of labels, and sufficiently small to penetrate the tissue section. The markers are localized by first introducing a biotinylated link into the tissue section and thereafter exposing the section to the labelling complex.
Inventors
- 👤 Su-Ming Hsu
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 22:56
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/581; G01N2474/00; G01N2474/20; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/962; Y10S435/975
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
103 words
A method for localizing markers on a prepared tissue section, said method comprising: exposing the tissue section to a primary reagent capable of specifically binding to the marker; exposing the tissue section to biotinylated antibody raised against the primary reagent; exposing the tissue section to a preformed labelling complex formed by reacting avidin and a biotinylated macromolecules, wherein at least some of said biotinylated macromolecules are detectable enzymes and said avidin and said biotinylated macromolecules are present at a weight ratio in the range from 1:1 to 16:1; and localizing the marker by reacting the detectable enzyme with substrate and observing the reaction.
Claim 2
Independent Claim
103 words
A method for localizing markers on a prepared tissue section, said method comprising: exposing the tissue section to a primary reagent capable of specifically binding to the marker; exposing the tissue section to biotinylated antibody raised against the primary reagent; exposing the tissue section to a preformed labelling complex formed by reacting avidin and a biotinylated macromolecules, wherein at least some of said biotinylated macromolecules are detectable enzymes and said avidin and said biotinylated macromolecules are present at a weight ratio in the range from 1:1 to 16:1; and localizing the marker by reacting the detectable enzyme with substrate and observing the reaction.
Claim 3
Independent Claim
86 words
A method for localizing markers on a prepared tissue section, said method comprising: introducing a biotinylated reagent into the tissue section, said reagent capable of binding to the tissue section substantially at the marker site; exposing the tissue section to the labelling complex comprised of avidin which has been reacted with a preselected amount of a biotinylated enzyme, wherein the weight ratio of avidin to the biotinylated enzyme, reacted was in the range from 1:1 to 16:1, respectively; and detecting the enzyme by reaction with substrate.
Claim 4
Independent Claim
86 words
A method for localizing markers on a prepared tissue section, said method comprising: introducing a biotinylated reagent into the tissue section, said reagent capable of binding to the tissue section substantially at the marker site; exposing the tissue section to the labelling complex comprised of avidin which has been reacted with a preselected amount of a biotinylated enzyme, wherein the weight ratio of avidin to the biotinylated enzyme, reacted was in the range from 1:1 to 16:1, respectively; and detecting the enzyme by reaction with substrate.
Claim 5
Dependent Claim
51 words
A labelling complex for binding to a biotinylated receptor introduced onto a biological specimen, said complex comprising avidin which has been reacted with a preselected amount of a biotinylated enzyme, wherein the weight ratio of avidin to the biotinylated enzyme reacted was in the range from a 1:6 to 16:1, respectively.
Claim 6
Dependent Claim
51 words
A labelling complex for binding to a biotinylated receptor introduced onto a biological specimen, said complex comprising avidin which has been reacted with a preselected amount of a biotinylated enzyme, wherein the weight ratio of avidin to the biotinylated enzyme reacted was in the range from a 1:6 to 16:1, respectively.
Claim 7
Dependent Claim
20 words
A labelling complex as in claim 3, wherein the avidin:biotinylated detectable enzyme is in the range from 2:1 to 4:1.
Claim 8
Dependent Claim
24 words
A labelling complex as in claim 3, wherein the biotinylated enzyme has a molecular weight in the range from 10,000 to 2×10 6 daltons.
Claim 9
Dependent Claim
24 words
A labelling complex as in claim 3, wherein the biotinylated enzyme has a molecular weight in the range from 25,000 to 6×10 5 daltons.
Claim 10
Dependent Claim
15 words
A labelling complex as in claim 3, wherein the biotinylated enzyme is biotinylated horseradish peroxidase.
Claim 11
Dependent Claim
75 words
A kit for preparing a labelling complex for binding to a biotinylated receptor, the kit comprising, a preselected quantity of avidin having a predetermined concentration; a preselected quantity of biotinylated detectable enzyme having a predetermined concentration; and instructions to admix the avidin and the biotinylated enzyme in a buffer solution in amounts so that the resulting weight ratio of avidin:biotinylated enzyme in the buffer prior to reaction is in the range from 1:1 to 16:1.
Claim 12
Dependent Claim
75 words
A kit for preparing a labelling complex for binding to a biotinylated receptor, the kit comprising, a preselected quantity of avidin having a predetermined concentration; a preselected quantity of biotinylated detectable enzyme having a predetermined concentration; and instructions to admix the avidin and the biotinylated enzyme in a buffer solution in amounts so that the resulting weight ratio of avidin:biotinylated enzyme in the buffer prior to reaction is in the range from 1:1 to 16:1.
Claim 13
Dependent Claim
28 words
A kit as in claim 8, wherein the instructions specify that the weight ratio of avidin:biotinylated enzyme to be admixed is in the range from 2:1 to 4:1.
References Cited (Backward Citations)
US4298685A
Cited
US4228237A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1990-08-03 | 1991-02-02 | 1991-08-04 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1994-08-03 | 1995-02-02 | 1995-08-04 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1998-08-03 | 1999-02-02 | 1999-08-04 | $7700.00 | Window Closed / Lapsed |