Brilliant Blue R250

• CAT Number: R017025
• CAS Number: 6104-59-2
• Molecular Formula: C45H44N3NaO7S2
• Molecular Weight: 825.97
• Purity: ≥95%
• Synonyms: sodium;3-[[4-[[4-(4-ethoxyanilino)phenyl]-[4-[ethyl-[(3-sulfonatophenyl)methyl]azaniumylidene]cyclohexa-2,5-dien-1-ylidene]methyl]-N-ethylanilino]methyl]benzenesulfonate
• InChI: InChI=1S/C45H45N3O7S2.Na/c1-4-47(31-33-9-7-11-43(29-33)56(49,50)51)40-23-15-36(16-24-40)45(35-13-19-38(20-14-35)46-39-21-27-42(28-22-39)55-6-3)37-17-25-41(26-18-37)48(5-2)32-34-10-8-12-44(30-34)57(52,53)54;/h7-30H,4-6,31-32H2,1-3H3,(H2,49,50,51,52,53,54);/q;+1/p-1
• InChIKey: NKLPQNGYXWVELD-UHFFFAOYSA-M
• SMILES: CCN(CC1=CC(=CC=C1)S(=O)(=O)[O-])C2=CC=C(C=C2)C(=C3C=CC(=[N+](CC)CC4=CC(=CC=C4)S(=O)(=O)[O-])C=C3)C5=CC=C(C=C5)NC6=CC=C(C=C6)OCC.[Na+]

Brilliant blue R250 (Brilliant Blue R), an anionic dye, is the most popular stain to detect proteins resolved in SDS-PAGE gels[1].
Stain solution preparation:
Add 100 mL of glacial acetic acid to 450 mL ultrapure water.
Dissolve the 3 g of Coomassie Dye in 450 mL methanol.
Filter the solution before use
Standard Gel Staining Protocol
1. Gel may be prefixed in 50% MeOH, 10% HoAC, and 40% H2O for 30 minutes to overnight.
2. Stain gel in the above solution, with 0.25-0.3% Coomassie Blue R-250, for 2-4 hours, until the gel is a uniform blue color. Staining is complete when the gel is no longer visible in the dye solution. Berore complete staining, the gel will appear as a lighter area against the dark staining solution.
3. Destain for 4-24 hours in 5% MeOH, 7.5% HOAC, 87.5% H2O.
4. Store gels in 7% HOAC.

Reference

[1]. Krause RGE, et al. Crystal violet stains proteins in SDS-PAGE gels and zymograms. Anal Biochem. 2019;566:107-115.
 [Content Brief]