Immunoprecipitation of Target Protein-DNA Complexes from Motor Neuron Lysate

Begin with a tube containing pre-treated magnetic beads coated with antibodies specific to a target protein cross-linked to the DNA.   

Add a motor neuron lysate containing DNA fragments and incubate with agitation.

The DNA fragments interact with antibody-coated beads, forming bead-DNA complexes.

Centrifuge to collect the liquid from the cap and place it on a magnetic separator to precipitate the bead-DNA complexes.

Remove the supernatant containing unbound DNA fragments.

Introduce a detergent-based lysis buffer to solubilize non-specifically bound DNA fragments.

Centrifuge and perform the magnetic separation, then remove the supernatant.

Wash with a high salt buffer to disrupt non-specific interactions. Centrifuge, perform the magnetic separation, and remove the supernatant.

Wash with LiCl buffer to remove the remaining non-specific interactions. Centrifuge, perform the magnetic separation, and remove the supernatant.

Finally, wash with Tris-HCl buffer, centrifuge, and perform the magnetic separation.

Remove the supernatant and collect the beads containing the target protein-DNA complexes.

To begin the chromatin immunoprecipitation, wash the antibody-coated beads in 1 milliliter