Dear all: Can I ask for some help-opinion on possible problems for a superdex200 column to separate a glutaraldehyde-crosslinked sample?
We are using 0.1% Glutaraldehyde to crosslink and protein complex and after incubation, we stop the reaction by adding 50mM Tris buffer. We have analyzed the sample on an SDS-PAGE gel and we do not observe aggregates, only the crosslinked complex and some uncrosslinked protein. We would like to further analyze this sample on a gel filtration Superdex 200 Increase 3.2/300 column and although I cannot see any incompatibility in the column specifications, I would like to have some advice on how you deal with this. Since the glutaraldehyde is quenched with Tris, I imagine that passing this sample through the column would not make any harm in future purifications or will have a bad effect on resin stability. Would I need to wash the column with something special? Do you have columns dedicated to glutaraldhyde-containing samples? I am a bit concerned on “contaminating” the column or losing resolution in the future. Many thanks in advance and all best wishes for 2022, María J. Sánchez-Barrena ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
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