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Epigenetics Forum: DNA Methylation, Histone and Chromatin Study An Epigenetics Forum Dealing with DNA Methylation, the study of histones and chromatin


ChIP Chromatin Immunoprecipitation Lysis Buffers?

ChIP Chromatin Immunoprecipitation Lysis Buffers? - Epigenetics Forum: DNA Methylation, Histone and Chromatin Study

ChIP Chromatin Immunoprecipitation Lysis Buffers? - An Epigenetics Forum Dealing with DNA Methylation, the study of histones and chromatin


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Old 02-26-2008, 08:16 AM
Pipette Filler
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Question ChIP Chromatin Immunoprecipitation Lysis Buffers?



Hi every1,
I have questions on cell lysis for ChIP.

After cross-linking my cells, what lysis buffer should I use?

In different protocols they use different lysis buffers. Some lyse their cells directly using 1% SDS-Lysis buffer but others lyse the cells and isolate the nuclei and afterwards lyse the nuclei.

Are there any reasons why?

Also which is better RIPA buffer or SDS-lysis buffer? Does it depend on the cell line? I am currently using transfected HEK 293 cells.

Thanks in advance
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Old 04-15-2008, 03:27 PM
Pipette Filler
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Default Re: ChIP Chromatin Immunoprecipitation Lysis Buffers?

Hi Dear,
Though I am new to this area but found very good concept about choosing the lysis buffer in abCam website. In short ,Ripa is more stringent than NP-40 or Triton-X100. It depends on your antigen. Read IP protocol in their website that will help you to make a decision.
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Old 04-15-2008, 08:39 PM
Pipette Filler
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Default Re: ChIP Chromatin Immunoprecipitation Lysis Buffers?

Thank you very much for your reply. i will read the protocol there.

thanks
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Old 01-21-2009, 09:53 AM
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Default Re: ChIP Chromatin Immunoprecipitation Lysis Buffers?

hi, i work on preimplantation embryos. I would like to use ChIP for the analyasis of DNA methylation.. can anyone suggest how many embryos I need or how much DNA I need to analyse global DNA methylation?

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buffers , chip , chromatin , immunoprecipitation , lysis


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