Hello,
I am looking to capture a GST tagged protein on a glass slide. I was wondering if anyone had any experiences with glutathoine agarose resin. I realize this is commonly used in affinity chromatography and can adhere to chromatography columns. Does anyone have any expierience with this resin, is agarose resin able to firmly attach to glass? Any methods available to improve attachment of the agarose?
For chromatography, the gel beads are not "attached" to the column, the column is simply filled with them, they are held in place by filter gauze at top and bottom of the column.
What you probably want is different: a thin, optically homogeneous, layer of modified agarose on your slide. The agarose in your beads is probably cross-linked, so you can't simply melt it (check the doku, though). So you need to get unmodified agarose, dissolve it in buffer by boiling it (try different concentration from 1-5%), dip the slide briefly into the solution, so that you get a thin film, and let it harden o/n in the fridge in a humid chamber. Then modify with glutathione, there are kits for doing that, you need a kit for coupling to the OH-groups of sugars, not to the amino-groups of another protein.
Dear Chris,
alternatively you can coat the glass with Maleimide PEG and then attach the glutathione. Or you can simply buy maleimide slides ready for glutathione attachment :)
http://www.proteinslides.com/order
Best of luck!
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