The Potential of Proteomics: A Clear Path to Discovery | Part II


Posted by Carolyn P on Feb 22, 2017 3:00:00 AM

See a real life example of how PTMScan technology can facilitate translational discovery. In this short video we describe how CST not only identified a major driver of NSCLC, which can respond to an FDA approved drug, but went a step further and developed an antibody that can be used to test which patients might be candidates for treatment. All thanks to the power of simplified proteomics.

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Topics: Proteomics, PTMScan, techniques

The Potential of Proteomics: A Clear Path to Discovery


Posted by Carolyn P on Jan 11, 2017 3:00:00 AM

Whether your lab is well-versed and equipped to do Mass Spec analysis in-house or your lab has never considered setting up a proteomics experiment before, the Proteomics group at CST has a solution to help clear the way.  Click below to watch the video and learn more.

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Topics: Proteomics, PTMScan, techniques

The Cost of a Failed Western Blot


Posted by Claire S on Sep 14, 2016 3:00:00 AM


Thinking about lab expenses isn’t as enjoyable as investigating your favorite signaling pathway. However, because research money is hard to come by, it is something that should be considered when picking your reagents for western blotting. It makes sense to keep the quality of your primary antibody in mind, because the success of the entire experiment depends on the antibody being reliable, specific, and sensitive. If the antibody does not perform as expected your experiment may fail . . . and the cost of a failed western blot may be more than you think.

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Topics: Antibody Performance, Antibody Validation, Western Blot, techniques, Reproducibility

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...

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

The Cost of a Failed Western Blot


Posted by Claire S on Aug 13, 2015 8:13:02 PM


Thinking about lab expenses isn’t as enjoyable as investigating your favorite signaling pathway. However, because research money is hard to come by, it is something that should be considered when picking your reagents for western blotting. It makes sense to keep the quality of your primary antibody in mind, because the success of the entire experiment depends on the antibody being reliable, specific, and sensitive. If the antibody does not perform as expected your experiment may fail . . . and the cost of a failed western blot may be more than you think.

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Topics: Antibody Performance, Antibody Validation, Western Blot, techniques

Intracellular Flow Cytometry in Action


Posted by Liana G on Aug 12, 2015 8:00:00 AM


Traditionally, flow cytometry has been used to identify distinct cell types within a heterogeneous pool of cells, based on extracellular or surface marker expression, an application commonly known as immuno-phenotyping. However, this technology is also readily amenable to intracellular target detection and can be successfully applied to the study of complex signaling events.

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Topics: Protocols, Flow, Cell Biology, techniques

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.

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Topics: Protocols, ChIP, techniques

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


Posted by Carolyn P on Jul 29, 2015 5:00:00 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, techniques

4 Steps to Better Immunohistochemistry: Steps 2, 3, and 4.


Posted by Carolyn P on Jul 15, 2015 7:00:00 AM

This is part two of a two-part series on how to optimize your IHC protocols. Part one introduced the principles behind antigen retrival. Click here if you missed it...but, if you've got your tissue prepped and ready to go, we'll move on to the next steps in the staining protocol.

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Topics: Protocols, IHC, techniques

4 Steps to Better Immunohistochemistry: Step 1 - Antigen Retrieval


Posted by Carolyn P on Jul 8, 2015 7:00:00 AM

It’s Friday night - you’re tucked away in a dark, little room filled with microscopes.You could be out with your friends right now, but you begged off because you were certain this immunohistochemistry was going to reveal some small - but important - mystery of the universe to you.

Instead, you’re sitting here, cursing said universe and all its inhabitants, because all you see when you stare down the barrel of the scope is some indistinct fuzziness. And did the controls work - meh - who’s to say? There’s no sugar coating it – it’s a fail.

So, what’s next? If you’re like me, you’ll wing the slide into the trash with as much gusto as you can muster and head out to find your friends. Tomorrow, you’ll re-evaluate the experiment.

But where do you start? You probably know that a highly specific, high-affinity primary antibody is key to a successful IHC. But did you know that the companion reagents (i.e., buffers, etc.), which establish the pH and ionic strength of the system, are just as important? These reagents can influence the binding of the primary antibody to its epitope and dramatically affect the outcome of the assay.

To help you pick the best reagents for your assay (and make sure those Friday nights in the lab are worth the effort) we will spend the next several posts reviewing how companion reagents affect IHC. And, as an example we will describe our experience optimizing the protocol for one of our antibodies, PLK1 (208G4) Rabbit mAb.

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Topics: Protocols, IHC, Companion Reagents, techniques

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