Chromatin Immunoprecipitation by Neus Visa & Antonio Jordán-Pla

Author:Neus Visa & Antonio Jordán-Pla
Language: eng
Format: epub
Publisher: Springer New York, New York, NY

4.Mouse, rabbit, and goat IgGs are not always suitable controls. We have found that some IgGs can be extremely noisy. A control IgG under stringent conditions should not precipitate any identifiable chromatin as assessed by polymerase chain reaction (PCR) amplification. If a control immunoglobulin is precipitating noticeable amounts of chromatin, other sources should be tested. For precipitation antibodies, one general rule of thumb is that an antibody that cannot efficiently immuno-deplete lysates of its target protein will not be suitable for ChIP. This may seem obvious but few experimentalists consider this before they spend long hours unsuccessfully precipitating the chromatin they desire. Another consideration, but not necessarily feasible in every situation, is that an antibody raised against a protein that directly binds DNA will likely be more efficient than an antibody directed to a protein that binds DNA indirectly or through an ancillary protein.

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