I am trying to elute proteins from magnetic beads by immobilizing biotinylated DNA to the beads and therefore elute proteins from the beads, but unfortunately after elution, I can't see any band. Here is my protocol !
Slurry-18 microliter( binds to 3000ng of DNA)
DNA=3500ng
Elution Buffer used=10mM Tris +0.6M NaCl
Protein=1ml( Concentration= 3400ng/microliter)
ANy suggestions would be deeply appreciated !
The first three wells are the ladder, DNA protein mix and the flowthrough. After that I can't see any band for the elutions or the washes.
http://www.merckmillipore.com/chemicals/en_US/Merck-International-Site/USD/ViewProductDocuments-File?ProductSKU=EMD_BIO-72190&DocumentType=USP&DocumentId=%2Femd%2Fbiosciences%2Fuserprotocols%2Fen-US%2FTB529.pdf&DocumentSource=GDS
It seems that nothing was adsorbed to the column. May be that high affinity proteins were bound therefore you should elute with higher salt concentrations.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- DNA