Kit for use in electrophoretic matrix staining
Staining compositions and methods for use with a matrix, such as an electrophoretic gel, containing separated biopolymers. The compositions including an acid, an organic solvent, and a generally planar, fluid-permeable gel contact sheet consisting primarily of a non-cellulosic material. The acid and organic solvent may be sorbed to the fluid-permeable gel contact sheet, or may be sorbed to a layers contactable with a non-gel contact side of the gel contact sheet. In one embodiment, the composition is a source electrophoretic stain composition including a staining reagent. In one embodiment, the composition is a sink electrophoretic stain composition including a sorbent.
1. A kit for electrophoretically staining a biopolymer in a matrix comprising
a first non-cellulosic or modified cellulosic matrix contact sheet contacting a first side of the matrix,
a second matrix contact sheet contacting a second side of the matrix and comprising a non-cellulosic or partially cellulosic material,
a stain solution comprising a stain reagent and a solvent, the stain solution optionally being absorbed within the first matrix contact sheet,
and
a destain solution comprising an organic solvent, an acid, water, and optionally a buffering agent, the destain solution being absorbed within the second matrix contact sheet.
2. The kit of claim 1 where the stain solution is provided absorbed into the first matrix contact sheet.
3. The kit of claim 1 where the stain solution comprises 25% ethanol, 5% acetic acid, and 0.01% Coomassie Brilliant Blue dye R-250 or G-250, and the destain solution comprises 25% ethanol and 5% acetic acid.
4. The kit of claim 1 further comprising instructions for staining the biopolymer in the matrix using an electric field.
5. The kit of claim 1 where the first matrix contact sheet further comprises at least one additional sheet distal from the first side of the matrix and/or the second matrix contact sheet further comprises at least one additional sheet distal from the second side of the matrix, the at least one additional sheet being the same or different from the first matrix contact sheet and/or the second matrix contact sheet.
6. The kit of claim 1 where the first matrix contact sheet is capable of absorbing at least 5 ml of the stain solution, but less than 15 ml of the stain solution; a thickness of the first matrix contact sheet is less than 0.5 inches; and/or the matrix contact surface is sufficient to provide maximum contact between the first matrix contact sheet and the matrix.
7. The kit of claim 1 where the first matrix contact sheet or the first and the second matrix contact sheet is polyester, polyester/cellulose blend, polypropylene, polypropylene/cellulose blend, polyacrylamide, nitrocellulose, non-cellulose, polysaccharide, agarose, felt, polyvinylidene fluoride (PVDF), polyvinylidene difluoride, fine woven cotton, wool, regenerated cellulose fiber, Lyocell, and combinations thereof.
8. The kit of claim 7 where the first matrix contact sheet is polyester, polyester/cellulose blend, polypropylene, polypropylene/cellulose blend, polyacrylamide, nitrocellulose, non-cellulose, polysaccharide, agarose, felt, polyvinylidene fluoride (PVDF), polyvinylidene difluoride, regenerated cellulose fiber, Lyocell, and combinations thereof.
9. The kit of claim 5 where the at least one additional sheet is polyester, polyester/cellulose blend, polyacrylamide, nitrocellulose, polysaccharide, agarose, felt, polyvinylidene fluoride (PVDF), polyvinylidene difluoride, a fabric, a foam, a sponge, and combinations thereof.
10. The kit of claim 5 where the stain solution is present in the at least one additional sheet which is not in contact with the matrix.
11. The kit of claim 1 where the second matrix contact sheet comprises a sorbent selected from the group consisting of wound dressing non-adhesive material, woven and non-woven cotton, foam, felt, agarose, agarose/polyacrylamide matrix, alumina, silica, polysaccharides, polymer, and combinations thereof.
12. The kit of claim 1 where the stain solution comprises an acid, an organic solvent, and the stain reagent.
13. The kit of claim 12 where the acid is acetic acid and the organic solvent is ethanol or isopropanol.
14. The kit of claim 12 where the stain solution further comprises a buffering agent.
15. The kit of claim 14 where the buffering agent is sodium phosphate.
16. The kit of claim 1 where the stain reagent is a protein stain, and is selected from the group consisting of Coomassie Blue R-250, Coomassie Blue G-250, silver stain, hydrophobic or charged staining reagents, neutral staining reagents, and protein modification stains.
17. The kit of claim 16 where the charged staining reagents include chromogenic, fluorescent, chemiluminescent, and/or bioluminescent molecules.
18. The kit of claim 16 where the neutral staining reagent further comprises a non-charged organic or non-organic solvent.
19. The kit of claim 16 where the protein modification is at least one of phosphorylation, glycosylation, acetylation, nitrosylation, SUMOylation, or epitope tags.
20. The kit of claim 1 where the matrix is selected from the group consisting of SDS polyacrylamide gels, native polyacrylamide gels, membranes, and filters.