Hello, Guys.
I'm planning to use Sepharose CL gel for my studies related to liposomes.
But, I have no idea how to pack empty PD-10 column (or even smaller) with the Sepharose gel.
In the manufacturer protocol (supplied by GE), it seems that requires the pump to fill columns with the gel. However, in the lab, we don't have those kinds of pumps to fill.
Therefore, I'd like to fill the gel with gravity or centrifugation.
If any of you have experience with Sepharose gel packing, could you give any protocol or advice about how to do that?
Best,
Normally after degassing the gel and buffer you can pack it in normal room temperature and pressure manually.
Dear Jeonghwan,
for our Compact Cartridges (Cube Biotech, 1 and 5 ml) we have designed an instruction with pictures, which unfortunately is not linked on our website in the moment. You can get it by writing a short mail to [email protected].
Here's the data sheet for our 1 ml compact cartridge with a short filling procedure (without pictures):
https://cube-biotech.com/media/pdf/40/1d/27/PureCube-Compact-Cartridge-1-mL.pdf
I have packed Sepharose S200, Sephadex G25, G75 and G100 gel at a larger scale for preparative fractionation. The method is to degas the gel and soak in buffer overnight until it swells fully and then pack in glass column under normal condition. It is important to pack slowly until the gel settles and leave it at least 24h after packing. The following day you can run the buffer and then start loading samples. This technique should work but you can do a trial and error. (note: if the gel is already in swollen state or in solution it may take less time to do the 1st step).
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