Chromatin Immunoprecipitation: Difference between revisions
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==== 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 | # Add 50 μl re-suspended magnetic bead slurry 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. | # Place the microfuge tubes on the magnet rack and remove supernatants. | ||
# Resuspend the beads in 1 ml cold PBS/BSA. | # Resuspend the beads in 1 ml cold PBS/BSA. | ||
# Repeat Steps b and c 3 times. | # Repeat Steps b and c 3 times. | ||
# Add 200 μl PBS/BSA to beads. | # Add 200 μl PBS/BSA to beads. | ||
# Add | # Add 1 μg primary antibody. Do not vortex beads after adding the antibody. | ||
# Gently mix on a rotator platform for at least 2 hours at 4°C. | # 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. | # Wash beads 3 times (steps b-c), resuspending the beads by inverting the tubes during each wash. | ||