162
edits
Changes
→Couple the primary antibody for each transcription factor or chromatin protein to magnetic beads
==== Couple the primary antibody for each transcription factor or chromatin protein to magnetic beads====
# Add 20 μl re-suspended magnetic bead slurry (for each sample you plan to use with this antibody) to a 1.5 ml microfuge tube on ice containing 1 ml cold PBS/BSA. Vortex briefly to mix well.
# Place the microfuge tubes on the magnet rack and remove supernatants.
# Resuspend the beads in 1 ml cold PBS/BSA.
# Repeat Steps b and c 3 times.
# Add 200 μl PBS/BSA (for each sample you plan to use this antibody with) to beads.# Add 1 μg primary antibody. Mix gently by tapping--Do not vortex beads after adding the antibody.
# Gently mix on a rotator platform for at least 2 hours at 4°C.
# Wash beads 3 times (steps b-c), resuspending the beads by inverting the tubes during each wash.
# Resuspend in 100 μl PBS/BSA(for each sample you plan to use this antibody with), and proceed to Step 2.
==== Incubate bead-antibody complex with fragmented, cross-linked chromatin====