US4555490A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Rapid visualization system for gel electrophoresis
Assignee: United States Department of Health and Human Services
•
Filing: 1984-06-08
•
Granted: 1985-11-26
•
Expires: 2004-06-08
Abstract
Abstract A method using light ("photodevelopment") to develop a metallic silver image of biopolymers, particularly nucleic acids and proteins separated on polyacrylamide gels, whereby it is possible to visualize protein and nucleic acid patterns within 10 minutes after electrophoretic separation. This "photodevelopment" method requires only two solutions: a solution to "fix" the proteins and a solution containing silver ions, which produces an image when exposed to light. This type of protein stain has achieved a sensitivity of about 0.5 ng of protein. DNA separated on polyacrylamide may also be visualized with this stain.
Inventors
- 👤 Carl R. Merril
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 14:33
CPC Classifications
G; G01; G01N; G01N27/00; G01N27/26; G01N27/416; G01N27/447; G01N27/44704; G01N27/44717; G01N27/44721; G01N27/44726; Y; Y10; Y10S; Y10S436/00; Y10S436/905; Y10T; Y10T436/00; Y10T436/14; Y10T436/142222; Y10T436/143333; Y10T436/25
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
117 words
In a method for separating biopolymer entities by thin membrane or polyacrylamide gel electrophoresis of the type including fixing the separated entities in a suitable fixing medium and thereafter rapidly rinsing the fixed gel with deionized water to remove the fixing medium, the improvement comprising having in said fixing medium about 2% w/v citric acid and about 0.2% w/v sodium chloride; and visualizing said separated entities in less than about 10 minutes by placing said gel or membrane in a single solution photoimaging mixture of 50% v/v methanol, 10% v/v acetic acid and 40% v/v deionized water containing 2% w/v silver nitrate and then uniformly illuminating said gel or membrane with visible light within any chemical treatment.
Claim 2
Dependent Claim
117 words
In a method for separating biopolymer entities by thin membrane or polyacrylamide gel electrophoresis of the type including fixing the separated entities in a suitable fixing medium and thereafter rapidly rinsing the fixed gel with deionized water to remove the fixing medium, the improvement comprising having in said fixing medium about 2% w/v citric acid and about 0.2% w/v sodium chloride; and visualizing said separated entities in less than about 10 minutes by placing said gel or membrane in a single solution photoimaging mixture of 50% v/v methanol, 10% v/v acetic acid and 40% v/v deionized water containing 2% w/v silver nitrate and then uniformly illuminating said gel or membrane with visible light within any chemical treatment.
Claim 3
Dependent Claim
14 words
The method of claim 1, wherein said biopolymer entities are proteins or nucleic acids.
Claim 4
Dependent Claim
133 words
A process for rapid visualization of separated biopolymers on suitable support medium comprising: (a) fixing the biopolymers separated on said support medium for about 5 min in a fixing medium consisting esentially of about 50% v/v methanol, about 10% v/v acetic acid and about 40% v/v of deionized water containing about 2% w/v citric acid and about 0.2% w/v sodium chloride; (b) thereafter rinsing said support medium with deionized water to remove surface chloride; (c) placing said support medium in a single solution photoimaging mixture containing about 50% v/v methanol, about 10% v/v acetic acid and 40% v/v of deionized water containing about 2% w/v silver nitrate; and (d) illuminating said support medium with uniform light without any chemical treatment, wherein imaging of separated biopolymers appears either in less than about 10 minutes.
Claim 5
Dependent Claim
133 words
A process for rapid visualization of separated biopolymers on suitable support medium comprising: (a) fixing the biopolymers separated on said support medium for about 5 min in a fixing medium consisting esentially of about 50% v/v methanol, about 10% v/v acetic acid and about 40% v/v of deionized water containing about 2% w/v citric acid and about 0.2% w/v sodium chloride; (b) thereafter rinsing said support medium with deionized water to remove surface chloride; (c) placing said support medium in a single solution photoimaging mixture containing about 50% v/v methanol, about 10% v/v acetic acid and 40% v/v of deionized water containing about 2% w/v silver nitrate; and (d) illuminating said support medium with uniform light without any chemical treatment, wherein imaging of separated biopolymers appears either in less than about 10 minutes.
Claim 6
Dependent Claim
13 words
The process of claim 3 wherein said support medium is polyacrylamide electrophoresis gel.
Claim 7
Dependent Claim
21 words
The process of claim 3 wherein said biopolymers are selected from the group consisting of proteins, nucleic acids and mixtures thereof.
Claim 8
Dependent Claim
13 words
The process of claim 3 wherein said support medium is an electrophoresis membrane.
Claim 9
Dependent Claim
13 words
The process of claim 6 wherein said membrane is a cellulose nitrate membrane.
Claim 10
Dependent Claim
119 words
A process for rapid visualization of separated biopolymers on suitable support medium comprising: (a) immersing said support medium for about 5 min in an aqueous solution consisting essentially of about 50% v/v methanol, about 10% v/v acetic acid and about 40% v/v deionized water containing about 2% w/v citric acid and about 0.2% w/v sodium chloride; (b) thereafter immediately rinsing said support medium with deionized water to remove surface chloride; (c) placing said support medium in a a single photoimaging mixture containing about 50% v/v methanol, about 10% v/v acetic acid and 40% v/v of deionized water containing about 2% w/v silver nitrate, wherein imaging of separated biopolymers appears immediately upon placing said support medium in said photoimaging mixture.
Claim 11
Dependent Claim
119 words
A process for rapid visualization of separated biopolymers on suitable support medium comprising: (a) immersing said support medium for about 5 min in an aqueous solution consisting essentially of about 50% v/v methanol, about 10% v/v acetic acid and about 40% v/v deionized water containing about 2% w/v citric acid and about 0.2% w/v sodium chloride; (b) thereafter immediately rinsing said support medium with deionized water to remove surface chloride; (c) placing said support medium in a a single photoimaging mixture containing about 50% v/v methanol, about 10% v/v acetic acid and 40% v/v of deionized water containing about 2% w/v silver nitrate, wherein imaging of separated biopolymers appears immediately upon placing said support medium in said photoimaging mixture.
Claim 12
Dependent Claim
13 words
The process of claim 8 wherein said support medium is an electrophoresis membrane.
Claim 13
Dependent Claim
13 words
The process of claim 9 wherein said membrane is a cellulose nitrate membrane.
References Cited (Backward Citations)
WO1982003128A1
Cited
US4405720A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1988-11-25 | 1989-05-27 | 1989-11-26 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-11-25 | 1993-05-27 | 1993-11-26 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-11-25 | 1997-05-27 | 1997-11-26 | $7700.00 | Window Closed / Lapsed |