Chromatin immunoprecipitation (ChIP) explained: principle and procedure | ChIP assay

This video lecture describes
1. What is chromatin ?
2. What is chromatin immunoprecipitation?
3. When and why Chromatin immunoprecipitation is performed?
4. What are the different steps involved in Chromatin Immunoprecipitation (ChIP) ?
5. How ChIP analysis is performed ?
6. What is Chromatin Immunoprecipitation protocol?
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[@alessiailas4929]

Thank you😭 that's the best explanation I found among all the RUclips videos on the subject

[@BiologyLectures]

You are most welcome.

[@alhaeri1]

thank you

[@BiologyLectures]

You are most welcome.

[@mohammadgufran9560]

Nice explanation

[@BiologyLectures]

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[@user-tv5sj7pt2c]

this is a fantastic lecture. i hope to do another for plant bioinformatic and biostatistics as must, i am from egypt

[@BiologyLectures]

Thank you very much. Please consider subscribing us. For bioinformatics lectures, please go through this playlist ruclips.net/p/PLi1EfMhc7RNr0XMmUCkKZWItx-TvBg0cY

[@investincrypto2859]

Very well explained about chip. Thanks a million

[@BiologyLectures]

Glad it was helpful!

[@acampinglamp]

Thanks for posting!

[@BiologyLectures]

You are most welcome

[@dikshyapanthi7681]

WOW best explanation 😊👍

[@BiologyLectures]

Thanks a lot 😊

[@anujvermabiotech]

Thanks

[@BiologyLectures]

Welcome

[@shawlineema7431]

extraordinary video!!

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[@dakshinapanthi722]

Excellent explanation 😊

[@BiologyLectures]

Thank you.

[@harshitashrivastava8877]

Great Explanation🙂

[@BiologyLectures]

Thank you very much

[@Hari788]

Excellent explanation. Thank you 😊

[@BiologyLectures]

You are welcome!

[@user-mz7rl3ub7y]

thx🥰

[@BiologyLectures]

you are most welcome

[@aishamughal3094]

Really well explained!

[@BiologyLectures]

Thank you very much 😊

[@grinsunsharma567]

great ...

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[@linaabdelghany4033]

Thank you for your explanation. May I ask a question
1- After the fixation step then use lysis buffer but how can we isolate the protein DNA complex from the cytoplasmic protein ? Do we use isolation kit or centrifuge?
Thanks

[@BiologyLectures]

Since DNA is present in the nucleus, the protein DNA interaction described in the video is also nuclear. In the method, described in the video, we will not have protein DNA complex in the cytoplasm. Hope this helps.

[@linaabdelghany4033]

@@BiologyLectures so as in the method the DNA is the one that will purified and analyzed in the end of the process, it should be fine if the antibody bind to other proteins ( non DNA bound).
Thank you for your clarification :)

[@BiologyLectures]

@@linaabdelghany4033 We use Chip grade antibody specific for the protein of interest. Lets say that if we want to study the interaction between protein X and the transcription factor Y. The Chip grade antibody used will be specific for protein X. If there is interaction between the protein of interest X transcription factors A, this complex will be immunoprecipitated by the specific antibody used. Its not at all fine if the antibody is not specific.
There are two major considerations when choosing an antibody for ChIP-whether it will work in ChIP and whether it is specific. Ideally, the target will have an antibody that has already been shown to work in ChIP or in another immunoprecipitation (IP) application.

[@linaabdelghany4033]

@@BiologyLectures Thank you very much for your explanation.

[@BiologyLectures]

@@linaabdelghany4033 you are most welcome

[@achanyasjayan9195]

How can we protect chromatin protein from the agents which is used to remove cytosolic proteins in cell lysis stage.

[@BiologyLectures]

Crosslinking help fix protein DNA complex Chromatin) stable. If sonication is used in lysis, keep the chromatin on ice and limit the pulse to no more than 30 seconds to avoid protein denaturation due to heat.

[@achanyasjayan9195]

@@BiologyLectures thankyou❤

[@BiologyLectures]

@@achanyasjayan9195 You are most welcome

[@zer.e.zehranaqvi6439]

how we can analyze in the last step that DNA and proteins are interacting or not ?

[@zer.e.zehranaqvi6439]

plz explain the last step in detailed

[@BiologyLectures]

We use the antibody specific for the protein in a DNA protein complex. And finally, we can analyze the data by performing qPCR. If we observe the Ct values for the gene of interest, that means that DNA and protein are interacting. We can also analyze by performing sequencing.

[@zer.e.zehranaqvi6439]

@@BiologyLectures Thank you

[@BiologyLectures]

@@zer.e.zehranaqvi6439 You are most welcome.