System for pH-neutral stable electrophoresis gel
View Patent ↗A gel and buffer system for gel electrophoresis wherein separation occurs at neutral pH.
1. A discontinuous buffer gel electrophoresis system comprising:
an electrophoresis slab gel comprising a separating gel having between 8 and 25 (wt/vol) % acrylamide, and a gel buffer having a pH between 6 and 8, wherein the gel buffer comprises bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane;
an anode buffer; and
a cathode buffer solution comprising 3-(N-morpholino)propanesulfonic acid (MOPS).
2. The discontinuous buffer gel electrophoresis system according to claim 1 , wherein the gel buffer comprises at least 0.3 molar bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane.
3. The discontinuous buffer gel electrophoresis system according to claim 1 , wherein the gel buffer comprises 0.3 to 0.4 molar bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane.
4. The discontinuous buffer gel electrophoresis system according to claim 1 , wherein the gel buffer comprises 0.357 molar bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane.
5. The discontinuous buffer gel electrophoresis system according to claim 1 , wherein the gel is polymerized using at least 5% crosslinker.
6. The discontinuous buffer gel electrophoresis system according to claim 1 , wherein the gel has a pH of between 6.5 and 7.5.
7. The discontinuous buffer gel electrophoresis system according to claim 1 , wherein the system further comprises a buffer comprising a reducing agent that is negatively charged at a neutral pH.
8. The discontinuous buffer gel electrophoresis system according to claim 1 , wherein the cathode buffer comprises a sodium or tris salt of 3-(N-morpholino)propanesulfonic acid (MOPS).
9. The discontinuous buffer gel electrophoresis system according to claim 1 , wherein the separating gel comprises sodium dodecyl sulfate.
10. The discontinuous buffer gel electrophoresis system according to claim 1 , wherein the gel is in a disposable cassette.
11. The discontinuous buffer gel electrophoresis system according to claim 1 , wherein the gel is enclosed in a sealed package, wherein the sealed package comprises the gel buffer.
12. The discontinuous buffer gel electrophoresis system according to claim 11 , wherein the gel is between 3 weeks and 1 year old.
13. The discontinuous buffer gel electrophoresis system according to claim 12 , wherein the gel has not been subjected to electrophoresis.
14. The discontinuous buffer gel electrophoresis system according to claim 11 , wherein the gel is between 2 months and 1 year old.
15. The discontinuous buffer gel electrophoresis system according to claim 11 , wherein the gel is at least 1 year old.
16. The discontinuous buffer gel electrophoresis system according to claim 11 , wherein the gel is 1 year old.
17. The discontinuous buffer gel electrophoresis system according to claim 11 , wherein the gel is at least 3 weeks old.
18. The discontinuous buffer gel electrophoresis system of claim 1 further comprising an electrophoresis unit.
19. The discontinuous buffer gel electrophoresis system of claim 1 , wherein the same gel buffer is present throughout the electrophoresis slab gel.
20. A method for performing gel electrophoresis, comprising performing electrophoresis to separate a protein using a discontinuous buffer gel electrophoresis system comprising:
an electrophoresis slab gel comprising a separating gel having between 8 and 25 (wt/vol) % acrylamide, and a gel buffer having a pH between 6 and 8, wherein the gel buffer comprises bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane;
an anode buffer; and
a cathode buffer solution comprising 3-(N-morpholino)propanesulfonic acid (MOPS).
21. The method of claim 20 , further comprising the following steps prior to performing electrophoresis:
(i) casting the electrophoresis slab gel and
(ii) storing the electrophoresis slab gel in the gel buffer after casting and before performing the electrophoresis.
22. The method of claim 21 , wherein the electrophoresis is performed between 2 months and 1 year after the gel has been cast and stored.
23. The method of claim 21 , wherein the electrophoresis is performed 1 year after the gel has been cast and stored.
24. The method of claim 21 , wherein the gel is cast by an outside vendor, before electrophoresis is performed.
25. The method of claim 24 , further comprising shipping the gel to a laboratory before performing electrophoresis and after the gel has been cast.
26. The method of claim 21 , wherein the gel is polymerized using 5% crosslinker.
27. The method of claim 21 , wherein the gel is cast in a disposable cassette.
28. The method of claim 21 , wherein the gel is cast with a gel buffer having a pH of between 6.5 and 7.5.
29. The method of claim 21 , wherein the electrophoresis is performed between 3 weeks and 1 year after the gel has been cast and stored.
30. The method of claim 20 , wherein the gel buffer comprises at least 0.3 molar bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane.
31. The method of claim 20 , wherein the gel buffer comprises between 0.3 and 0.4 molar bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane.
32. The method of claim 20 , wherein the gel buffer comprises between 0.35 and 0.36 molar bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane.
33. The method of claim 20 , wherein the gel is removed from a sealed package before performing electrophoresis.
34. The method of claim 20 , wherein the gel comprises sodium dodecyl sulfate.
35. The method of claim 20 , wherein the discontinuous buffer gel electrophoresis system further comprises a reducing agent, wherein the reducing agent migrates into the gel while performing the electrophoresis to separate the protein.
36. The method of claim 35 , wherein the reducing agent is sodium bisulfite or a negatively charged thiol.
37. The method of claim 35 , wherein the reducing agent is thioglycolic acid.
38. The method of claim 20 , wherein the cathode buffer comprises a sodium or tris salt of 3-(N-morpholino)propanesulfonic acid (MOPS).
39. The method of claim 20 , wherein the separating is carried out in a gel comprising between 8 and 20 (wt/vol) % acrylamide.
40. The method of claim 20 , wherein the separating is carried out in a gel comprising between 8 and 15 (wt/vol) % acrylamide.
41. The method of claim 20 , wherein the separating is carried out in a gel comprising between 8 and 12 (wt/vol) % acrylamide.
42. The method of claim 20 , wherein the separating is carried out in a gel comprising between 8 and 12 (wt/vol) % acrylamide to separate proteins between 2 and 200 kDa.
43. The method of claim 20 , wherein the same gel buffer is present throughout the electrophoresis slab gel.
44. The discontinuous buffer gel electrophoresis system according to claim 43 , wherein the separating gel comprises between 8 and 15 (wt/vol) % acrylamide.
45. The discontinuous buffer gel electrophoresis system according to claim 44 , wherein the separating gel comprises between 8and 12 (wt/vol) % acrylamide.
46. The discontinuous buffer gel electrophoresis system according to claim 43 , wherein the separating gel comprises between 8 and 15 (wt/vol) % acrylamide.
47. The discontinuous buffer gel electrophoresis system according to claim 46 , wherein the separating gel comprises between 8 and 12 (wt/vol) % acrylamide.
48. A discontinuous buffer gel electrophoresis system comprising:
an electrophoresis slab gel comprising a separating gel having between 8 and 25 (wt/vol) % acrylamide, and a gel buffer having a pH between 6 and 8, wherein the gel buffer is bis(2-hydroxyethyl)iminotris (hydroxymethyl)methane;
an anode buffer; and
a cathode buffer solution comprising 3-(N-morpholino)propanesulfonic acid (MOPS).
49. The discontinuous buffer gel electrophoresis system according to claim 48 , wherein the gel buffer comprises at least 0.3 molar bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane.
50. The discontinuous buffer gel electrophoresis system according to claim 48 , wherein the gel buffer comprises 0.3 to 0.4 molar bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane.
51. The discontinuous buffer gel electrophoresis system according to claim 48 , wherein the gel buffer comprises 0.357 molar bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane.
52. The discontinuous buffer gel electrophoresis system according to claim 48 , wherein the gel is polymerized using at least 5% crosslinker.
53. The discontinuous buffer gel electrophoresis system according to claim 48 , wherein the gel has a pH of between 6.5 and 7.5.
54. The discontinuous buffer gel electrophoresis system according to claim 48 , wherein the system further comprises a buffer comprising a reducing agent that is negatively charged at a neutral pH.
55. The discontinuous buffer gel electrophoresis system according to claim 48 , wherein the cathode buffer comprises a sodium or tris salt of 3-(N-morpholino)propanesulfonic acid (MOPS).
56. The discontinuous buffer gel electrophoresis system according to claim 48 , wherein the separating gel comprises sodium dodecyl sulfate.
57. The discontinuous buffer gel electrophoresis system according to claim 48 , wherein the gel is in a disposable cassette.
58. The discontinuous buffer gel electrophoresis system of claim 48 , wherein the same gel buffer is present throughout the electrophoresis slab gel.
59. The discontinuous buffer gel electrophoresis system of claim 48 further comprising an electrophoresis unit.
60. The discontinuous buffer gel electrophoresis system according to claim 48 , wherein the gel is enclosed in a sealed package, wherein the sealed package comprises the gel buffer.
61. The discontinuous buffer gel electrophoresis system according to claim 60 , wherein the gel is at least 3 weeks old.
62. The discontinuous buffer gel electrophoresis system according to claim 60 , wherein the gel is between 3 weeks and 1 year old.
63. The discontinuous buffer gel electrophoresis system according to claim 60 , wherein the gel has not been subjected to electrophoresis.
64. The discontinuous buffer gel electrophoresis system according to claim 60 , wherein the gel is between 2 months and 1 year old.
65. The discontinuous buffer gel electrophoresis system according to claim 60 , wherein the gel is at least 1 year old.
66. The discontinuous buffer gel electrophoresis system according to claim 60 , wherein the gel is 1 year old.