Dimethyl pimelimidate dihydrochloride

Guidelines (The following is our recommended protocol, which serves only as a guideline and should be modified according to your specific needs)^[1].
An alternative fixation protocol using Dimethyl pimelimidate dihydrochloride.
Experimental Procedures:
1. Remove a vial of Dimethyl pimelimidate dihydrochloride from the refrigerator, allow it to warm to room temperature before opening (this substance is highly hygroscopic).
2. Gently rinse cells with PBS, then fix the cells directly in the culture dish or transfer them to a 1.5 mL centrifuge tube, followed by shaking for fixation at room temperature for 1 hour (use 5 mL Dimethyl pimelimidate dihydrochloride/DMSO/PBS for a 10 cm culture dish, and 400 μL Dimethyl pimelimidate dihydrochloride/DMSO/PBS for a 1.5 mL centrifuge tube).
3. Add 1% paraformaldehyde (dissolved in PBS) directly to the well plate/tube to achieve a final paraformaldehyde concentration of 2/3% (for example, add 10 mL of 1% paraformaldehyde to 5 mL of DMP solution, or add 800 μL of paraformaldehyde to 400 μL of solution).
4. Seal the plate with Parafilm, or incubate the samples in 1.5 mL tubes with rotation at room temperature in a fume hood for 5-20 hours (the incubation time is determined based on experimental results for different proteins).
5. Centrifuge at 364 × g for 5 minutes, then rinse the cells with PBS, taking care not to leave sample residues on the tube wall.
6. Add lysis buffer and initiate the ChIP assay. Alternatively, the pellet can be stored frozen at this point.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.