Protein market pellets and method of making and using the same

• US 20030196897A1
• Filed: 04/15/2003
• Published: 10/23/2003
• Est. Priority Date: 04/19/2002
• Status: Active Grant

• Current Assignee: Expedeon Incorporated (2invest AG)
• Original Assignee: Pagegel Incorporated
• Inventors: Alpenfels, William F., Vilalta, Patricia Marilu
• Granted Patent: US 7,153,403 B2
• Time in Patent Office: Days
• US Class Current: 204/461
• CPC Class Codes: G01N 27/44726 using specific dyes, markers or binding molecules
• CPC Class Codes: Y10T 436/105831 Protein or peptide standard or control [e.g., hemoglobin, etc.]

Abstract

A protein marker pellet for use in a gel electrophoresis system includes one or more protein markers including a protein of known molecular weight with ionic components to provide electrical conductance, and a non-protein, gel-forming material having a porosity sufficient to allow the one or more protein markers to migrate out of the protein marker pellet under the influence of an electrical field at the same rate as the ionic components.

Claims

1. A protein marker pellet for use in a gel electrophoresis system, comprising:

one or more protein markers including a protein of known molecular weight with ionic components to provide electrical conductance;

a non-protein, gel-forming material having a porosity sufficient to allow the one or more protein markers to migrate out of the protein marker pellet under the influence of an electrical field at the same rate as the ionic components.

2. The protein marker pellet of claim 1, wherein the protein marker pellet is at least one of a solid and a semi-solid.

3. The protein marker pellet of claim 1, wherein the gel electrophoresis system is a SDS PAGE gel electrophoresis system.

4. The protein marker pellet of claim 1, wherein the gel-forming material is a thermal-setting polymer.

5. The protein marker pellet of claim 4, wherein the thermal-setting polymer is high-melt agarose.

6. The protein marker pellet of claim 5, wherein the high-melt agarose includes an agarose concentration between 0.5 and 2%.

7. The protein marker pellet of claim 6, wherein the agarose concentration is about 1%.

8. The protein marker pellet of claim 1, wherein the one or more protein markers include multiple protein markers with proteins of different sizes.

9. The protein marker pellet of claim 1, wherein the protein marker pellet includes SDS.

10. The protein marker pellet of claim 1, wherein the protein marker pellet includes buffer salts compatible with the gel electrophoresis system.

11. The protein marker pellet of claim 1, wherein the protein marker pellet includes a density agent.

12. The protein marker pellet of claim 11, wherein the density agent is at least one of glycerol and di-methyl-formamide.

13. The protein maker pellet of claim 1, further including a substrate that the protein marker pellet is fixed to.

14. The protein marker pellet of claim 13, wherein the gel electrophoresis system includes a reservoir with multiple wells having a length and a width, and the substrate includes a length and a width substantially the same as the length and the width of the multiple wells.

15. A method of making a protein marker pellet for use in a gel electrophoresis system, comprising:

providing a protein marker solution including one or more protein markers having a protein of known molecular weight with ionic components to provide electrical conductance;

providing a non-protein, gel-forming solution;

warming the protein marker solution and the gel-forming solution;

combining the protein marker solution and the gel-forming solution to form a protein marker gel solution;

providing the protein marker gel solution into multiple small volumes of protein marker gel solution;

allowing the multiple small volumes of protein marker gel solution to set so as to form multiple protein marker pellets having a porosity sufficient to allow the protein marker to migrate out of the protein marker pellet under the influence of an electrical field at the same rate as the ionic components.

16. The protein marker pellet of claim 15, wherein the gel electrophoresis system is a SDS PAGE gel electrophoresis system.

17. The method of claim 15, wherein the protein marker pellets are at least one of a solid and a semi-solid.

18. The method of claim 15, wherein the gel-forming material is a thermal-setting polymer.

19. The method of claim 18, wherein the thermal-setting polymer is high-melt agarose.

20. The method of claim 19, wherein the high-melt agarose includes an agarose concentration between 0.5 and 2%.

21. The method of claim 20, wherein the agarose concentration is about 1%.

22. The method of claim 15, wherein the one or more protein markers include multiple protein markers with proteins of different sizes.

23. The method of claim 15, wherein the protein marker pellet includes SDS.

24. The method of claim 15, wherein the protein marker pellet includes buffer salts compatible with the gel electrophoresis system.

25. The method of claim 15, wherein the protein marker pellet includes a density agent.

26. The method of claim 25, wherein the density agent is at least one of glycerol and di-methyl-formamide.

27. The method of claim 15, further including dropping the protein marker gel solution into multiple small volumes of protein marker gel solution onto a substrate.

28. The method of claim 27, wherein the substrate is paper.

29. The method of claim 28, wherein the paper is a lacquer-coated paper.

30. The method of claim 27, wherein the substrate is plastic film.

31. The method of claim 27, wherein the substrate includes perforations.

32. The method of claim 27, wherein the gel electrophoresis system includes a reservoir with multiple wells having a length and a width, and the substrate includes multiple strips of a length and a width substantially the same as the length and the width of the multiple wells.

33. The method of claim 15, wherein warming the protein marker solution includes warming the protein marker solution to about 50 degrees C.

34. The method of claim 15, wherein warming the gel-forming solution includes warming the gel-forming solution to about 50 degrees C.

35. The method of claim 15, wherein combining the protein marker solution and the gel-forming solution includes combining the protein marker solution with the gel-forming solution in substantially a 3:

2 ratio.

36. The method of claim 15, wherein the small volumes of protein marker gel solution are 3 to 10 microliters.

37. A method of using a protein marker pellet with a gel electrophoresis system including multiple wells and an electrophoresis gel, comprising:

providing a protein marker pellet including one or more protein markers having a protein of known molecular weight with ionic components to provide electrical conductance, and a non-protein, gel-forming material having a porosity sufficient to allow the protein marker to migrate out of the protein marker pellet under the influence of an electrical field at the same rate as the ionic components;

depositing the protein marker pellet in one of the wells of the gel electrophoresis system;

causing the protein marker to migrate out of the protein marker pellet under the influence of an electrical field.

38. The method of claim 37, wherein the protein marker pellet is at least one of a solid and a semi-solid.

40. The method of claim 37, wherein the gel-forming material is a thermal-setting polymer.

41. The method of claim 40, wherein the thermal-setting polymer is high-melt agarose.

42. The method of claim 41, wherein the high-melt agarose includes an agarose concentration between 0.5 and 2%.

43. The method of claim 42, wherein the agarose concentration is about 1%.

44. The method of claim 37, wherein the one or more protein markers include multiple protein markers with proteins of different sizes.

45. The method of claim 37, wherein the protein marker pellet includes SDS.

46. The method of claim 37, wherein the protein marker pellet includes buffer salts compatible with the gel electrophoresis system.

47. The method of claim 37, wherein the protein marker pellet includes a density agent.

48. The method of claim 47, wherein the density agent is at least one of glycerol and di-methyl-formamide.

49. The method of claim 37, further including a substrate that the protein marker pellet is fixed to, and depositing the protein marker pellet includes depositing the protein marker pellet in one of the wells onto an electrophoresis gel with the protein mark pellet faced down.

50. The method of claim 49, wherein the gel electrophoresis system includes a reservoir with multiple wells having a length and a width, and the substrate includes a length and a width substantially the same as the length and the width of the multiple wells.