Dear all,
I am cross-linking cells with formaldehyde and then i would like to extract DNA which is cross-linked with the proteins and then from this DNA Cross-linked protein complex, I would like to extract all bound proteins (reverse cross-linking) to send it for Mass spectrometry. I want to extract all of the cross-linked proteins not a specific one.
Do you have any suggestion how can i achieve it?
Do you think using Ampure XP beads it is possible ?
or may be first precipitating DNA with ethanol and then extracting proteins from that precipitate using SDS sample buffer followed by heating at 95 degrees for 25 mins and loading it directly to gel.
I appreciate your time and response.
Kind regards
Niharika
You have given a great thought. It's fine to precipitate DNA with chilled ethanol, collect previpitates and digest with low conc (1 mg/mL) of DNAse. Protein shall survive and after passing it through a Sephadex G-10 column use protein fraction for analysis.
Dear Shamsher Singh Kanwar,
Thank you for your response and your time. I will try this method out.
Kind regards
NIharika
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts