Method and apparatus for extraction of species from electrophoresis media

• US 4,699,706 A
• Filed: 05/30/1986
• Issued: 10/13/1987
• Est. Priority Date: 05/30/1986
• Status: Expired due to Fees

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Bio-Rad Laboratories Incorporated
• Inventors: Chu, Daniel Y., Burd, Samuel
• Primary Examiner(s): Niebling, John F.
• Assistant Examiner(s): Starsiak, Jr., John S.
• Application Number:
  US06/868,964
• Time in Patent Office: 501 Days
• Field of Search: 204/180.1, 204/182.3, 204/299 R, 204/301, 204/182.8, 204/182.6, 204/182.1, 204/182.7, 204/415, 204/416, 204/417, 204/418, 204/419, 204/420, 210/445, 210/446, 210/232, 210/927
• US Class Current: 204/613
• CPC Class Codes: G01N 27/4473 by electric means

Abstract

Electrophoretic extraction of molecular species from a separation medium such as a gel segment is achieved by placing the separation medium in a tube closed at one end by a retaining material such as glass frit, and sealing the closed end of the tube against the mouth of a receiving cup which has a semi-permeable membrane (such as a dialysis membrane) for its bottom. The tube with receiving cup attached is inserted in a tube electrophoresis cell such as those commonly used in analytical laboratories. As current passes through the tube, the molecular species migrates from the separation medium to pass through the retaining frit and collect on the membrane for subsequent recovery.

Claims

1. A receiving cup for collecting species electrophoretically extracted from a support medium, said receiving cup comprising:

a sleeve containing a transverse internal shoulder extending the full circumference thereof;

a membrane impermeable to said species, sized to rest on said shoulder;

a gasket of resilient material sized to fit snugly inside said sleeve and to rest on said shoulder above said membrane;

a ring of substantially nondeformable material sized to fit snugly and completely within said sleeve and to rest on said gasket; and

a protrusion on the interior wall of said sleeve of sufficient size to retain said ring, said protrusion positioned to compress said ring, said gasket and said membrane together between itself and said shoulder.

2. A receiving cup in accordance with claim 1 in which said protrusion is a transverse rib extending the full circumference of said sleeve.

3. Apparatus for separating species from a support medium during electrophoretic migration of said species, said apparatus comprising:

a tube closed at one end with a first material permeable to said species and impermeable to said support medium;

a vessel open at the top and closed at the bottom with a second material permeable to water and impermeable to said species;

a sleeve having first and second open ends sized to receive respectively said closed end of said tube and said top of said vessel;

a first rib on the interior wall of said sleeve to engage the exterior of said tube; and

a second rib on the interior wall of said sleeve to engage the exterior of the side wall of said vessel.

4. Apparatus for separating species from a support medium during electrophoretic migration of said species, said apparatus comprising:

a tube closed at one end with a first material permeable to said species and impermeable to said support medium;

a vessel open at the top and closed at the bottom with a second material permeable to water and impermeable to said species; and

a sleeve having first and second open ends sized to receive respectively said closed end of said tube and said top of said vessel, and an internal shoulder to abut the inserted end of one of said tube and said vessel.

5. Apparatus for separating species from a support medium during electrophoretic migration of said species, said apparatus comprising:

a tube closed at one end with a first material permeable to said species and impermeable to said support medium;

a vessel open at the top and closed at the bottom with a second material permeable to water and impermeable to said species, said tube and said vessel being of different outer diameters; and

a sleeve having first and second open ends sized to receive respectively said closed end of said tube and said top of said vessel, and an internal shoulder to abut the inserted end of the larger diameter one of said tube and said vessel.

6. Apparatus for separating species from a support medium during electrophoretic migration of said species, said apparatus comprising:

a tube closed at one end with a first material permeable to said species and impermeable to said support medium;

a vessel open at the top and closed at the bottom with a second material permeable to water and impermeable to said species;

a sleeve having first and second open ends sized to receive respectively said closed end of said tube and said top of said vessel; and

a flange extending laterally from the bottom of said vessel into said sleeve.

7. Apparatus for separating species from a support medium during electrophoretic migration of said species, said apparatus comprising:

(a) a tube closed at one end with a first material permeable to said species and impermeable to said support medium;

(b) a receiving cup comprising;

(i) a first sleeve containing a transverse internal shoulder extending the full circumference thereof;

(ii) a membrane impermeable to said species, sized to rest on said shoulder;

(iii) a gasket of resilient material sized to fit snugly inside said sleeve and to rest on said shoulder above said membrane;

(iv) a ring of substantially nondeformable material sized to fit snugly within said sleeve and to rest on said gasket; and

(v) a protrusion on the interior wall of said sleeve of sufficient size to retain said ring, said protrusion positioned to compress said ring, said gasket and said membrane together between itself and said shoulder; and

(c) a second sleeve having first and second open ends sized respectively to receive said closed end of said tube and said first sleeve.