Electrophoresis apparatus

• US 4,915,811 A
• Filed: 06/17/1988
• Issued: 04/10/1990
• Est. Priority Date: 06/19/1987
• Status: Expired due to Term

• Current Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Original Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Inventors: Yamamoto, Masayoshi, Yamamoto, Shoichi, Fujita, Akira
• Primary Examiner(s): Niebling, John F.
• Assistant Examiner(s): Starsiak, John S.
• Application Number:
  US07/208,399
• Time in Patent Office: 662 Days
• Field of Search: 204/299 R, 204/182.8, 204/182.9
• US Class Current: 204/619
• CPC Class Codes: G01N 27/44756 Apparatus specially adapted therefor
• CPC Class Codes: G01N 27/44773 Multi-stage electrophoresis, e.g. two-dimensional electrophoresis
• CPC Class Codes: G01N 27/44782 of a plurality of samples

Abstract

An electrophoresis apparatus comprises a gel sheet device composed of spacers disposed between two films at side edges thereof, a gel membrane provided within a space formed between the two films by the spacers, and sample-pouring portions provided near an upper edge of the gel membrane. Two supporting members hold the gel sheet device therebetween, and a gap-forming member contacts edges of at least one surface of the gel sheet device. Each sample-pouring portion is adapted to sample pouring into a space formed by the films, a shark'"'"'s teeth comb, and the upper edge face of the gel membrane. The gap-forming member remote from an upper buffer solution vessel is provided such that an inner edge of a gap-forming member portion contacting the upper portion of the gel sheet device coincides with the upper edge face of the gel membrane, or the extent of overlapping of the gap-forming member portion upon the gel membrane is 5 mm or less.

Claims

1. In an electrophoresis apparatus comprising two sheet members formed of a non-conductive organic polymer film and disposed to stand facing each other, spacers having predetermined thicknesses disposed at right and left edge portions between the two sheet members, and an eletrophoresis gel membrane of uniform thickness grasped between the two sheet members thereby forming an electrophoresis sheet, a pair of flat plate-like supporting members for supporting the electrophoresis sheet by sandwiching it between the supporting members, a frame-like spacer disposed between the electrophoresis sheet and one of the supporting members, and a flat plate-like comb member having teeth-like protrusions inserted in the top edge of the gel membrane to form sample liquid pouring apertures disposed along the membrane and defined by the top edge of the membrane, the upper edges of the spaced-apart sheet members, and the adjacent faces of the teeth-like protrusions, wherein the improvement comprises:

said frame-like spacer having an upper outer edge and an upper inner edge spaced downwardly from said upper outer edge to define the upper width of said frame, said frame being positioned with its upper inner edge substantially aligned with the upper edge of said gel membrane when said frame is in place.

2. An apparatus as defined in claim 1 wherein the upper edge of said gel membrane is spaced below the upper edge of one of said spaced-apart sheet members by a distance substantially equal to the upper width of said frame.

6. An apparatus as defined in claim 1, 2, 3, 4 or 5 wherein said comb is formed of a polyethylene terephthalate film.

7. An apparatus as defined in claim 1, 2, 3, 4 or 5 wherein said comb is formed of a semitransparent polyethylene terephtalate film.

8. An apparatus as defined in claim 1, 2, 3, 4 or 5 wherein said electrophoresis gel consists of polyacrylamide.

9. An apparatus as defined in claim 1, 2, 3, 4 or 5 wherein said spacers disposed between said sheet members have a thickness greater than the corresponding thickness of said gel membrane, and the thickness d of said comb is greater than the corresponding thickness of said spacers by 20 μ

m or less.

10. An apparatus as defined in claim 1, 2, 3, 4 or 5 wherein the thickness d of said comb and the thickness G of said gel membrane satisfy the relationship of

11. An apparatus as defined in claim 1, 2, 3, 4 or 5 wherein said frame-like spacer member is formed of an electrically insulating flexible thin sheet.

12. An apparatus as defined in claim 1, 2, 3, 4 or 5 wherein the thickness of said frame-like spacer member is within the range of 0.1 mm to 0.6 mm.

3. In an electrophoresis apparatus comprising two sheet members formed of a non-conductive organic polymer film and disposed to stand facing each other, spacers having predetermined thicknesses disposed at right and left edge portions between the two sheet members, and an eletrophoresis gel membrane of uniform thickness grasped between the two sheet members to form an electrophoresis sheet, a pair of flat plate-like supporting members for supporting the electrophoresis sheet by sandwiching it between the supporting members, a frame-like spacer disposed between the electrophoresis sheet and one of the supporting members, and a flat plate-like comb member having teeth-like protrusions inserted in the top edge of the gel membrane to form sample liquid pouring apertures disposed along the membrane and defined by the top edge of the membrane, the upper edges of the spaced-apart sheet members, and the adjacent faces of the teeth-like protrusions, wherein the improvement comprises:

said frame-like spacer having an upper outer edge and an upper inner edge spaced downwardly from said upper outer edge to define the upper width of said frame, a portion of said upper width overlapping an upper portion of said gel membrane by an amount no more than 5 mm along the longitudinal extent of said membrane.

4. An apparatus as defined in claim 3 wherein the extent of overlapping of said frame width relative to said gel membrane is 3 mm or less along the longitudinal extent of said membrane.

5. In an electrophoresis apparatus comprising two sheet members formed of a non-conductive organic polymer film and disposed to stand facing each other, spacers having predetermined thicknesses disposed at right and left edge portions between the two sheet members, and an eletrophoresis gel membrane of uniform thickness grasped between the two sheet members to form an electrophoresis sheet, a pair of flat plate-like supporting members for supporting the electrophoresis sheet by sandwiching it between the supporting members, a frame-like spacer disposed between the electrophoresis sheet and one of the supporting members, and a flat plate-like comb member having teeth-like protrusions inserted in the top edge of the gel membrane to form sample liquid pouring apertures disposed along the membrane and defined by the top edge of the membrane, the upper edges of the spaced-apart sheet members, and the adjacent faces of the teeth-like protrusions, wherein the improvement comprises:

said frame-like spacer having marking indicia adapted to determine the position at which said comb is to be inserted downwardly in said gel membrane, and

said frame-like spacer having an upper outer edge and an upper inner edge spaced downwardly from said upper outer edge to define the upper width of said frame, said upper inner edge of said frame and the upper edge of said membrane being in a fixed position and in sufficiently close proximity to each other such that said comb can be repeatedly accurately positioned in said gel membrane to prevent sample liquid poured in one aperture from invading an adjacent pouring aperture and to prevent upward swelling of said gel membrane during electrophoresis.