I am trying to reproduce an old purification protocol from 2004 that claims the use of a pseudo-affinity chromatography based on Green A Dyematrex gel (Amicon). The product is discontinued from Amicon catalog. Any alternatives around ?. Thanks
Hi! Beth from Practichem here, and I am sorry to hear that your reagent-in-need is no longer produced. The dye ligand in Green A Dyematrex also goes by Green HE4B (Procion) and Reactive Green 19 (Sigma), but Sigma has also decided to discontinue their Reactive Green 19 resin. The dye itself is available from several suppliers if you wanted to make your own resin by cross-linking the dye to agarose or sepharose, but that is a messy endeavor.
A Nature Protocols article discussed the use of Green A Dyematrex in matrix metalloproteinase purification, and acknowledged that it is no longer in production (doi:10.1038/nprot.2008.74). The authors recommended using a chromatography resin that had the dye ligand Cibacron Blue 3GA or Reactive Red 120. Attached is the Sigma product documentation for their Reactive Dye affinity chromatography matrices, which contains the chemical structures for all products. It would be wise to try both resins since the protein-ligand interactions for triazinyl dyes can be unique for each protein. A cursory comparison suggests that Reactive Red 120 may be a more conservative choice based on the chemical groups and linker length, however, Cibacron Blue could be just as effective as a ligand and the blue dye resins are much more common and possibly more cost effective in the long run.
Prometric makes a series of dye-ligand resins in its Mimetic series including one called Mimetic Green 1, however, I have not yet been able to determine the chemical structure of the dye. I have written the company to ask for product information and whether the ligand is similar to Reactive Green 19 or if it based off of a chelated metal center like Reactive Green 5. I would advise you to stick to the previously mentioned affinity resins until I can determine the nature of Mimetic Green 1.
Please let me know if you have any more questions, and if my advice was helpful. Good luck!
Thanks Elizabeth for the detailed answer. I will give a try for Cibacrom blue and Reactive red to see what happens.
I will let you know how it works in my hands. Best
Paco
Mimetic Blue 2 A6XL is the nearest equivalent product to Matrex Gel Green A available from ProMetic ([email protected])
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