Immunoprecipitation (IP) Troubleshooting Guide

Immunoprecipitation / IP troubleshooting involves adaptations of the general protocol, including optimization of buffer composition, appropriate volume of sample and buffers to use, as well as the length of time for incubation and the number of washing steps, etc. Many of the components of the Sino Biological immunoprecipitation KIT are also available separately.
The main challenge is to determine the appropriate conditions for the specific binding between the antigen that the antibody pulls down and its interacting partner(s). Polyclonal antibodies often perform better in IPs than monoclonal antibodies. Don't hesitate to test different antibodies to find the best one for your specific IP.

The lysis step
The binding step
The washing step
The elution step
The detection step

The lysis step
The binding step
The washing step
The elution step
The detection step

Antibody immobilized on magnetic beads.
Detection reagent specific to native antibody.

Set positive/negtive control
Check your buffers
Check your lysate