Patent US4920059A successfully refreshed from Google Patents.
US4920059A
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Staining method for proteins and nucleic acids
Assignee: Janssen Pharmaceutica NV
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Filing: 1985-06-12
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Granted: 1990-04-24
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Expires: 2005-06-12
Abstract
Abstract A process for staining proteins and nucleic acids wherein the stain is a suspension of colloidal metal particles and the proteins and nucleic acids are visualized as a colored signal localized at the binding site of the colloidal metal particles to the proteins or nucleic acids or quantitatively determined at this site following art-known spectrophotometric procedures.
Inventors
- 👤 Marc K. J. J. Moeremans
- 👤 Guido F. T. Daneels
- 👤 Marc C. De Raeymaeker
- 👤 Jan R. De Mey
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 07:12
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/68; G01N33/6803; G01N33/6827; G01N33/683; C; C12; C12Q; C12Q1/00; C12Q1/68; G01N27/00; G01N27/26; G01N27/416; G01N27/447; G01N27/44704; G01N27/44717; G01N27/44721; G01N27/44726; G01N33/6836; Y; Y10; Y10T; Y10T436/00; Y10T436/14; Y10T436/142222; Y10T436/143333
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
105 words
A process for staining nucleic acids in or on a solid support which comprises: (i) applying to nucleic acids which are in or on a solid support a suspension consisting essentially of colloidal metal particles, the concentration and pH of such suspension being chosen so as to provide full color saturation within an incubation time of up to one day; (ii) allowing the colloidal metal particles to bind to the nucleic acids thereby forming stained nucleic acids having colloidal metal particles attached to binding sites thereon; and (iii) visualizing or quantitatively determining said stained nucleic acids as a colored signal localized at said binding sites.
Claim 2
Dependent Claim
105 words
A process for staining nucleic acids in or on a solid support which comprises: (i) applying to nucleic acids which are in or on a solid support a suspension consisting essentially of colloidal metal particles, the concentration and pH of such suspension being chosen so as to provide full color saturation within an incubation time of up to one day; (ii) allowing the colloidal metal particles to bind to the nucleic acids thereby forming stained nucleic acids having colloidal metal particles attached to binding sites thereon; and (iii) visualizing or quantitatively determining said stained nucleic acids as a colored signal localized at said binding sites.
Claim 3
Dependent Claim
133 words
A process for staining proteins or nucleic acids fixed on an immobilizing matrix comprising the steps of: (i) immobilizing the proteins or nucleic acids to be detected on an immobilizing matrix; (ii) contacting the immobilizing matrix with a staining suspension consisting essentially of a sol of colloidal metal particles adjusted to a concentration and pH so as to provide full color saturation within an incubation time of up to one day, and supplemented with at least one detergent which promotes the specific binding of the colloidal metal particles to the proteins or nucleic acids, thereby forming stained proteins or stained nucleic acids having cololidal metal particles attached to binding sites thereon; and (iii) visualizing or qunatitatively determining said stained proteins or stained nucleic acids as a colored signal localized at said bonding sites.
Claim 4
Dependent Claim
133 words
A process for staining proteins or nucleic acids fixed on an immobilizing matrix comprising the steps of: (i) immobilizing the proteins or nucleic acids to be detected on an immobilizing matrix; (ii) contacting the immobilizing matrix with a staining suspension consisting essentially of a sol of colloidal metal particles adjusted to a concentration and pH so as to provide full color saturation within an incubation time of up to one day, and supplemented with at least one detergent which promotes the specific binding of the colloidal metal particles to the proteins or nucleic acids, thereby forming stained proteins or stained nucleic acids having cololidal metal particles attached to binding sites thereon; and (iii) visualizing or qunatitatively determining said stained proteins or stained nucleic acids as a colored signal localized at said bonding sites.
Claim 5
Dependent Claim
19 words
The process according to claim 2 wherein the immobilizing metrix is washed after each of steps i and ii.
Claim 6
Dependent Claim
27 words
The process according to claim 2 wherein the immobiliziation stap is effected by directly absorbing and/or covalently binding the proteins or nucleic acids to the immobilizing matrix.
Claim 7
Dependent Claim
78 words
The process according to claim 2 wherein the immobilizing matrix is washed after the immobilizing stap by contacting it with a buffer solution or water supplemented with at least one detergent which promotes staining specificity with staining specificity being defined as specific binding of the colloidal metal or metal compound particles to the proteins or nucleic acids and the absence of such binding on those parts of the immobilizing matrix where no protiens or nucleic acids are immobilized.
Claim 8
Dependent Claim
38 words
The process according to claim 5 wherein the at least one detergent in the buffer solution or water is selected from the group consisting of polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan mono-oleate, octyl phenoxy polyepoxy ethanol, and myristyltrimethylammonium bromide.
Claim 9
Dependent Claim
34 words
The process of claim 2 wherein the pH of such suspension is chosen so as to most closely result in said proteins or nucleic acids and said colloidal metal particles having opposite net charges.
Claim 10
Dependent Claim
121 words
A process for staining proteins or nucleic acids in a gel support comprising the steps of: (i) contacting a gel support supporting proteins or nucleic acids with a staininq suspension consisting essentially of colloidal metal particles adjusted to a concentration and pH which provides full color saturation within an incubation time of up to one day, thus forming stained proteins or stained nucleic acids having colloidal metal particles attached to binding sites thereon; and (ii) visualizing or quantitatively determining said stained proteins or stained nucleic acids as a colored signal localized at said binding sites, wherein the stained suspension is supplemented with at least one detergent which promotes specific binding of the colloidal metal particles to the proteins or nucleic acids.
Claim 11
Dependent Claim
121 words
A process for staining proteins or nucleic acids in a gel support comprising the steps of: (i) contacting a gel support supporting proteins or nucleic acids with a staininq suspension consisting essentially of colloidal metal particles adjusted to a concentration and pH which provides full color saturation within an incubation time of up to one day, thus forming stained proteins or stained nucleic acids having colloidal metal particles attached to binding sites thereon; and (ii) visualizing or quantitatively determining said stained proteins or stained nucleic acids as a colored signal localized at said binding sites, wherein the stained suspension is supplemented with at least one detergent which promotes specific binding of the colloidal metal particles to the proteins or nucleic acids.
Claim 12
Dependent Claim
75 words
The process according according to claim 8 wherein the gel support is washed after step i by contacting it with a buffer solution or water supplemented with at least one detergent which promotes staining specificity, with staining specificity being defined as specific binding of the colloidal metal particles to the proteins or nucleic acids and the absence of such binding on those parts of the gel support where no proteins or nucleic acids are located.
Claim 13
Dependent Claim
38 words
The process according to claim 9 wherein the at least one detergent in the buffer solution or water is selected from the group consisting of polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan mono-oleate, octyl phenoxy polyepoxy ethanol, and myristryltrimethylammonium bromide.
Claim 14
Dependent Claim
34 words
The process of claim 8 wherein the pH of such suspension is chosen so as to most closely result in said proteins or nucleic acids and said colloidal metal particles having opposite net charges.
Claim 15
Dependent Claim
32 words
The process according to one of claims 1, 2, or 8 wherein the colloidal metal particles consist of gold, silver or platinum metal or of gold, silver, platinum, iron or copper compounds.
Claim 16
Dependent Claim
36 words
The process according to one of claims 1, 2, or 8 wherein the colloidal metal particles consist of gold or silver metal or iron hydroxide amd have a particle size comprised between 1 and 100 nm.
Claim 17
Dependent Claim
66 words
The process according to one of claims 1, 2, or 8 wherein the suspension of colloidal metal particles is a colloidal suspension of iron hydroxide and the visualization of quantitative determination of the stained proteins or stained nucleic acids is effected after hydrolizing the iron hydroxide with an acid to form a ferric salt and treating the thus formed ferric salt with a ferrocyanide salt complex.
Claim 18
Dependent Claim
123 words
A process for staining proteins or nucleic acids in or on a solid support which comprises: (i) applying to proteins or nucleic acids which are in or on a solid solid support a suspension consisting essentially of colloidal metal particles other than colloidal silver particles, the concentration and pH of such suspension being chosen so as to provide full color saturation within an incubation time of up to one day; (ii) allowing the colloidal metal particles to bind to the proteins or nucleic acids thereby forming stained proteins or stained nucleic acids having colloidal metal particles attached to binding sites thereon; and (iii) visualizing or quantitatively determining said stained proteins or stained nucleic acids as a colored signal localized at said binding sites.
Claim 19
Dependent Claim
123 words
A process for staining proteins or nucleic acids in or on a solid support which comprises: (i) applying to proteins or nucleic acids which are in or on a solid solid support a suspension consisting essentially of colloidal metal particles other than colloidal silver particles, the concentration and pH of such suspension being chosen so as to provide full color saturation within an incubation time of up to one day; (ii) allowing the colloidal metal particles to bind to the proteins or nucleic acids thereby forming stained proteins or stained nucleic acids having colloidal metal particles attached to binding sites thereon; and (iii) visualizing or quantitatively determining said stained proteins or stained nucleic acids as a colored signal localized at said binding sites.
Claim 20
Dependent Claim
29 words
The process according to claim 15 wherein the colloidal metal particles consist of gold metal or iron hydroxide and have a particle size comprised between 1 and 100 nm.
Claim 21
Dependent Claim
34 words
The process of claim 15 wherein the pH of such suspension is chosen so as to most closely result in said proteins or nucleic acids and said colloidal metal particles having opposite net charges.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1993-04-23 | 1993-10-23 | 1994-04-24 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-04-23 | 1997-10-23 | 1998-04-24 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-04-23 | 2001-10-23 | 2002-04-24 | $7700.00 | Window Closed / Lapsed |