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. 

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

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Topics: ChIP, Tech Tips, Protocols, Primary Antibodies

Fluorescent Staining Using Multiple Antibodies


Posted by Tara W on Sep 13, 2017 3:00:00 AM


The use of multiple antibodies in a single experiment can provide useful information to researchers. Co-staining with multiple antibodies and cellular dyes is a simple, low-content form of multiplex analysis. Techniques for performing multiplex analyses in cells and tissues are powerful research tools that are applicable to general cell biology studies as well as diagnostic purposes. These techniques allow researchers to detect multiple biomarkers to assess their samples. They also allow for easy colocalization studies to determine relationships between analytes. Here we describe two common techniques for fluorescent staining using multiple antibodies in the same assay.

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Topics: IF-IC, techniques, Protocols, Companion Reagents, Primary Antibodies, Antibody Performance

Have You Ever Wondered: What's The Lowdown on Phospho-Specific Antibodies?


Posted by Ken B on Jul 26, 2017 3:20:00 AM

Early exploration of unmapped biological signaling pathways were carried out using radiolabeled phospho-imaging. The development of phospho-specific antibodies to detect and quantify protein phosphorylation made life easier for researchers (less 32P waste to deal with), but the interpretation of data from these experiments comes with its own set of caveats. 

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Topics: Western Blot, Antibody Performance, Post Translational Modification, techniques, Primary Antibodies

Successful Immunofluorescence: Antibody Dilution and Incubation Conditions


Posted by Ken B on May 10, 2017 3:01:00 AM

Part four of a series on immunofluorescence techniques. Check out previous posts on Validation, Experimental Controls, and Fixation/Permabilization

After your samples have been prepared, it's time to incubate them with a well-validated antibody,  the workhorse of immunofluorescence. If you are a seasoned pro at IF experiments, you are probably used to checking the antibody datasheet (or web page) for the recommended dilution. But have you ever wondered where those recommendations come from?
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Topics: IF-IC, techniques, Antibody Performance, Primary Antibodies, Protocols

Successful Immunofluorescence: Experimental Controls


Posted by Ken B on Mar 8, 2017 3:00:00 AM

 Part two of a four-part series on Immunofluorescence. Check out our posts on Validation and Fixation and Permeabilization

After months of hard work, your research has honed in on a hypothesis you can test with immunofluorescence (IF). You've chosen antibodies and performed pilot IF experiments (see The Importance of Validation), and the localization of the protein appears reasonable. But how can you be sure the IF data you've acquired represents real biological phenomena? We present two examples of experimental controls in this post.

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Topics: IF-IC, Primary Antibodies, Antibody Validation, Antibody Performance, Protocols, Post Translational Modification, Reproducibility

Successful Immunofluorescence: The Importance of Validation


Posted by Ken B on Mar 1, 2017 3:00:00 AM

Part one of a four-part series on Immunofluorescence. Check out Experimental Controls and Fixation and Permeabilzation.

After months of hard work, your research has zeroed in on a hypothesis you can test with immunofluorescence (IF). But now you have to make a choice. How do you decide which antibody to use to get reliable IF results? How do you know if the images are accurately reporting the target's localization? We explore some considerations in this post.

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Topics: IF-IC, Primary Antibodies, Antibody Validation, Reproducility, Antibody Performance, Protocols

Simplifying Proteomics - Part 2


Posted by Claire S on Feb 8, 2017 10:40:01 AM

Part 1 gave an overview on mass spectrometry-based proteomics. Now it’s time to talk about how this strategy can be used to identify peptides with post-translational modifications (PTM) from a complex biological sample.

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Topics: Primary Antibodies, Post Translational Modification, Proteomics

Why High Concentration doesn't always mean good Antibody Performance.


Posted by Carolyn P on Dec 14, 2016 3:00:00 AM

If you buy primary antibodies from different places, you may have noticed that many companies list the size of their products in micrograms per ml (i.e., by weight). You may have also noticed that CST does not - CST antibodies are listed by performance (e.g., 10 western blots) instead.

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Topics: Antibody Performance, Primary Antibodies

3 Questions to ask before starting your next immunofluorescence experiment.


Posted by Liana G on Oct 19, 2016 3:00:00 AM

Specificity, consistency, and optimized assay conditions are three key elements that help ensure reliable immunofluorescence (IF) staining results each and every time. So, before starting an experiment you should ask yourself the following three questions:

  • Is your antibody specific?
  • Is your antibody supported by an optimized IF protocol?
  • Is your antibody performing consistently?

Let's take them each in turn...

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Topics: Protocols, Primary Antibodies, Antibody Validation, IF-IC, Cell Biology

3 Questions to ask before starting your next immunofluorescence experiment.


Posted by Carolyn P on Sep 7, 2016 3:00:00 AM

Specificity, consistency, and optimized assay conditions are three key elements that help ensure reliable immunofluorescence (IF) staining results each and every time. So, before starting an experiment you should ask yourself the following three questions:

  • Is your antibody specific?
  • Is your antibody supported by an optimized IF protocol?
  • Is your antibody performing consistently?

Let's take them each in turn...

READ MORE >

Topics: Protocols, Primary Antibodies, Antibody Validation, IF-IC, Cell Biology, techniques

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