I found this video (silica gel was used): https://www.youtube.com/watch?v=8Ff1Xx-On-g&t=96s
Can I use this process to pack my sephadex onto the column? (yes, right after I swell sephadex).
Reena- That will work, although I tended not to put the sand on the top of the column. I didn't do that because the sand tends to fall into the Sephadex reducing the ability to reuse the material, if desired. I always lost a little Sephadex because of the sand at the bottom getting mixed with the resin. If you can use a column with a glass frit at the bottom, you don't need to use the sand. When running Sephadex, don't use air pressure to run the column faster as you might with silica. Gravity feed only!! Applying pressure causes the Sephadex to compress, which increases the pressure, causing it to compress...until the column can't run anymore.
Going beyond your question, and beyond LH-20, the video showed how to pack a column for loading a sample dissolved in solvent. You should consider looking up "silica column solid load"- you will get hits like this: https://chemtips.wordpress.com/2013/01/29/dry-loading-in-flash-chromatography/
Solid loading often gives better results, although you don't do it for Sephadex.
Fill the column reservoir to the top with solvent. Seal, attach to a pump, and open the column outlet. Pack at 300 cm/h until the bed has reached a constant height. Stop the flow, empty, and remove the packing reservoir. https://www.sigmaaldrich.com/technical-documents/protocols/biology/size-exclusion-chromatography/performing-a-separation-with-sephadex-lh-20.html
Kindly look at this discussion which had be done on research gate.
https://www.researchgate.net/post/How-do-I-pack-a-column-with-sephadex-LH-20
I normally pack it by MeOH first. Just soak new or used Sephadex in a beaker for hours, stir and pour into a column (I normally use 1.5cm*150cm column for separate natural products). Then use the eluent you designed (could be acetone mixed with water) to condition the column, I will never use a single solvent as eluent because I will change the ratio to control the separation. Don't forget to wash the column by MeOH after use. The Sephadex, therefore, is stored in the column with MeOH waiting for the next use. After around three times use you have to repack it.
Kindly visit..
https://www.sigmaaldrich.com/technical-documents/protocols/biology/size-exclusion-chromatography/performing-a-separation-with-sephadex-lh-20.html
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