I have seen it posted a couple of times that SDS must be diluted to 0.1% or less prior to ChIP. Why is that and is there a way to get rid of it other than dilution? Thx!
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Hi,
That is true since most antibodies used for IP are sensitive to high SDS concentration. You can do one of three things:
1. prepare nuclei, and shear in a non-ionic detergent shearing buffer
2. Titrate the SDS content of your shearing buffer to see how low you can go without effecting the shearing.
3. Dialyze your sample after shearing to get rid of SDS prior to IP.
Thank you
Hamid -
Thanks--I may try those! Although, do you think it would be bad to dilute my sample at least 1:10 to get SDS below 0.1%.Comment
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Not if you remain within the IP volume of the protocol you are following. Everyone using 1% SDS buffer for shearing, dilutes in the ChIP buffer for the IP step anyways.
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