Hi all,
I am using NAP-5 disposable columns from GE for removing DTT from my protein.
Can I use the same column again for the purpose of removing DTT.
Can you please mention the protocol for the reuse of NAP-5 columns?
Also in which buffer should I store the column until next use? (20%EtOH?)
Thanks a lot.
Of course. All you need to do is wash out the small molecules: Flush with 5 or more column volumes of water or PBS, then re-equilibrate with your sample buffer. For storage, add some biozide (sodium azide will do) and avoid letting them dry out. Too much ethanol will kill the sephahex resin by making the beads collapse.
The disposable NAP DNA purification columns is prepacked with Sephadex G-25 DNA Grade for gravity purification of nucleic acids (≥ 10mers). Surely you can reuse this column a number of times after after eluting DNA and washing the colum with 5-6 times the bed volume. You can store the colum matrix in 20% ethanol or 0.02% sodium azIde.
yes, i agree. U need to was them extensively. Best with the buffer u want to use next. We did that several times. Use 0,01% azide to prevent microbial growth. Or use 0,05% thiomersal, if u want to use it in a bio-assay....
PS: If u want to do it cheap in a small volume: use a bit glass-wool in a 1 ml pipette tip. fill it with 0,5 ml rehydrated G-25 sephadex (fine or superfine for protein; or larger for DNA). Then u can purify in a superlow volume. needs a bit optimization first (collect in small volume in a 96 well plate and then u know where ur stuff comes; use the perfect volumes later to isolate and wash)
Note that Cytiva (GE) has the PD series of desalting columns also. I believe they are cheaper than the NAP columns which are DNA-grade. Both can be reused as mentioned. Depending on how important cross contamination is to what you do you may want to CIP them between samples.
- Badges
- Science method