When I use it several times the efficiency of the purification of my protein is not as much good as usually.
Thanks for your advices
Since PD10 columns are just Sephadex G25 Med in a convenient column, the protocol for cleaning Sephadex G25 can allow you to reuse them multiple times.
See page 7 here:
https://www.gelifesciences.com/gehcls_images/GELS/Related%20Content/Files/1314716762536/litdoc56119096_20130410154337.pdf
transferring the resin to a buchner funnel can help in removing protein aggregates which tend to collect at the top of the column, as back flushing isn't possible otherwise. If you remember to be very gentle with the resin it can be reused many times.
PD10 can be used many times even without cleaning. Generally I monitor the flow rate and when the column starts to perform too slowly then I will replace it. I only use this procedure for a buffer exchange step rather than a purification as such. Also I only use the same PD10 with the same protein so no 'contamination'.
One thing that does hurt it is if you put pressure through the column ie wanting to push out the solution faster. Probably caused by a packing of the resin bed due to the increase in pressure, as afterwards the flow rates are slower and it takes longer to pass your solution through. Still works okay but takes longer to do.
Hi Natalia, PD10 has a long time of live without repacking. Only you must be careful and keep the column in good conditions. For example: samples and buffers must be filtered, avoid samples high viscosity. leave the column in some bacteriostatic (sodium azide, thiomersal)
I agree with the answers above.
In my experience, the key is to be gentle with PD10 columns. As mentioned earlier and in the GE handbook, it helps to :
a) avoid using different protein samples to avoid cross contamination
b) storing the column in a bacteriostatic buffer at the right temperature
c) ensuring the rate of resin packing & sample loading is appropriate (not too fast)
d) ensure the viscosity, pH, clarity of the loading solution is appropriate
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