Rapid visualization system for gel electrophoresis

• US 4,555,490 A
• Filed: 06/08/1984
• Issued: 11/26/1985
• Est. Priority Date: 06/08/1984
• Status: Expired due to Fees

• Current Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Original Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Inventors: Merril, Carl R.
• Primary Examiner(s): Richman, Barry S.
• Assistant Examiner(s): Hill, Jr., Robert J.
• Application Number:
  US06/618,949
• Time in Patent Office: 536 Days
• Field of Search: 436/86, 436/87, 436/88, 436/94, 436/164, 436/169, 436/515, 436/905, 436/174, 436/175, 436/176, 430/346, 430/616
• US Class Current: 436/86
• CPC Class Codes: G01N 27/44726 using specific dyes, markers or binding molecules
• CPC Class Codes: Y10S 436/905 Photochemical activation of reactions
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/25 including sample preparation

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.

Claims

1. 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.

2. The method of claim 1, wherein said biopolymer entities are proteins or nucleic acids.

3. 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.

4. The process of claim 3 wherein said support medium is polyacrylamide electrophoresis gel.

5. The process of claim 3 wherein said biopolymers are selected from the group consisting of proteins, nucleic acids and mixtures thereof.

6. The process of claim 3 wherein said support medium is an electrophoresis membrane.

7. The process of claim 6 wherein said membrane is a cellulose nitrate membrane.

8. 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.

9. The process of claim 8 wherein said support medium is an electrophoresis membrane.

10. The process of claim 9 wherein said membrane is a cellulose nitrate membrane.