Video: How to Choose Enzymatic or Sonication Protocol for ChIP


Posted by Ken B on Nov 6, 2019 3:00:00 AM

The success of your chromatin immunoprecipitation (ChIP) experiment depends on the fragmentation of chromatin, a critical step in the ChIP protocol. This can be accomplished with either sonication or enzymatic digestion. But how do you decide which chromatin fragmentation protocol to use in your ChIP experiments? A number of factors can influence your choice, making a decision seem daunting. So let’s simplify – watch the video and we'll will show you the way!

 

READ MORE >

Topics: Protocols, ChIP, techniques

Preparing a ChIP-seq DNA library. Where do I start?


Posted by Tamar A. on Sep 18, 2019 3:15:00 AM

Chromatin immunoprecipitation sequencing (ChIP-seq) is a flexible and powerful technique used by researchers to elucidate how gene regulation is involved with different biological events and with the progression of various conditions like cancer and neurodegenerative diseases.

READ MORE >

Topics: ChIP, Epigenetics

How to Choose Enzymatic or Sonication Protocol for ChIP


Posted by Chris S on Jun 26, 2019 3:10:00 AM

Researchers use chromatin immunoprecipitation, or ChIP, to identify and characterize protein-DNA interactions in the context of chromatin. ChIP experiments can use varying input samples, chromatin fragmentation methods, and provide ChIP-qPCR or ChIP-seq readouts.

READ MORE >

Topics: ChIP, Epigenetics

Tech Tips Video: How much antibody should I use for ChIP?


Posted by Ken B on Nov 28, 2018 3:15:00 AM

It's time to check out another video from the CST Tech Tips playlist! In this edition of Tech Tips, we'll tackle a common protocol question customers ask our ChIP team: how much antibody to use for chromatin immunoprecipitaion (ChIP) experiments. Adding more antibody isn't always better - watch the video to learn why. 

Can't see the embedded video above? Click the link below to play: 

How much antibody should I use in ChIP assays? | CST Tech Tips
 

Don't forget to subscribe to CST's YouTube channel for more videos to help your research!

 Subscribe

READ MORE >

Topics: Protocols, Primary Antibodies, ChIP, Tech Tips

Myth-busting for ChIP/ChIP-seq: Mono vs. Poly


Posted by Neha G and Ken B on Sep 26, 2018 3:15:00 AM

Researchers who run a lot of chromatin immunoprecipitation "ChIP" assays – maybe even your advisor – might subscribe to the idea that polyclonal antibodies perform better than monoclonal antibodies. But is that always actually true?

It’s worth your time to understand the differences between the two in terms of antigen recognition and specificity, and dispel some myths. 

READ MORE >

Topics: Antibody Performance, ChIP, Antibody Validation, Post Translational Modification, Reproducibility

Keep ChIP’ing away.


Posted by Curtis D on Jan 31, 2018 6:00:00 AM

So your experiments and data are funneling you down an inescapable path. You need to show direct gene regulation by your protein of interest. You think to yourself, “Oh, ChIP...”

READ MORE >

Topics: Protocols, Antibody Performance, ChIP, techniques, Reproducility

Need Better ChIP results? Use Enzymatic Digestion


Posted by Carolyn P on Aug 5, 2015 12:33:00 PM

The success or failure of a chromatin immunprecipitation (ChIP) experiment is highly dependent on the integrity of both the chromatin and the protein epitope, so your method of chromatin preparation can have a significant impact on the quality of your results.

READ MORE >

Topics: Protocols, ChIP, techniques

4 Steps to Better ChIP results - Step 4 - Downstream Analysis


Posted by Carolyn P on Mar 11, 2015 3:00:00 AM

Part one of this series described the importance of including proper controls to your protocols. In Part 2 we discussed how chromatin preparation affects the final outcome of the experiment. Then we spent a post considering the immunoprecipitating antibody - and now let's analyze the data.

READ MORE >

Topics: Protocols, ChIP, techniques

4 Steps to Better ChIP results - Step 3- Choosing an Antibody


Posted by Carolyn P on Mar 4, 2015 3:00:00 AM

Part one of this series described the importance of including proper controls to your protocols. In Part 2 we discussed how chromatin preparation affects the final outcome of the experiment. Now, let's take a minute to consider the immunoprecipitating antibody.

Antibodies that are not highly specific to the target of interest may bind unpredictably and increase the background noise; and this may make it more difficult to detect less abundant or lower stability interactions.

READ MORE >

Topics: Protocols, ChIP, techniques

4 Steps to Better ChIP results - Step 2- Prep well


Posted by Carolyn P on Feb 25, 2015 3:00:00 AM

Part one of this series described the importance of including proper controls to your protocols. Now, we will examine how chromatin preparation affects the final outcome of the experiment.

First consider the type of interaction you are trying to detect:

  • High-frequency, very stable protein-DNA interactions like those between histones and DNA, occur frequently enough that they may still be detected even if the protocol is not fully optimized.
  • Low-frequency, less stable interactions like the binding of polycomb group proteins to specific genes (e.g., Ezh2), may fall under the detection threshold if the protocol fails to safeguard the integrity of the protein and the DNA, or if it relies on an antibody that is not highly specific to the target of interest.

Now it's time to consider your options for preparing your chromatin...

READ MORE >

Topics: Protocols, ChIP, techniques

Subscribe to Email Updates

Recent Posts