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!

 

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Topics: Protocols, ChIP, techniques

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
 

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Topics: Protocols, Primary Antibodies, ChIP, Tech Tips

Video: Understanding Macrophages in the Tumor Microenvironment


Posted by Ken B on Sep 19, 2018 3:15:00 AM

Research trends in macrophage plasticity

It’s an exciting time for immuno-oncology research, as potential predictive biomarkers from an expanding collection of cell types are being pursued. Explore the plasticity of tumor-associated macrophages (TAMs) and challenges in distinguishing M1- versus M2- functional states in this 5-minute video featuring CST Developmental Scientist Sarah Klein, PhD. 

Can't see the embedded video? Click here to watch.

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Topics: Cancer Research, Immunology, Immunotherapy

Tech Tips Video: Milk or BSA? Choosing a blocking protein for Western Blot


Posted by Ken B on Sep 5, 2018 3:00:00 AM

So you've set the timer to five minutes for the first of three TBST washes for your western blot membrane. Now what? Sure, you could check your email or social media for the 30th time before lunch. Or you could do something informative, like check out a CST Tech Tips video! This is a new short video series featuring the same scientists who develop and validate CST antibodies, here to offer insights and protocol tips.

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Topics: Protocols, Western Blot, techniques, Tech Tips

Q&A: From Signaling Pathways to Career Pathways


Posted by Ken B on Aug 15, 2018 3:10:00 AM

Each summer, college and high school interns at CST get a chance to sample work life in a biotech environment. They are typically in the early stages of considering potential career paths, so what better time to set up a Q&A panel? Five CST employees shared some of their career development experiences during a recent “lunch and learn” session with the interns. Selections from the session have been transcribed and edited for brevity. –ed.

Q: Which do you like better, being on the bench or managing?

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Topics: Career Development

Have you ever wondered: Am I setting up my loading controls correctly?


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

You’re gathering data from all your experiments and preparing to present to your advisor and thesis committee at your annual progress report. You have an interesting hypothesis, and you have a validated antibody that recognizes your target protein on a western blot (WB). The molecular weight of the band is correct, and the expression of the target protein changes just the way you predicted it would. Now, you know — and you’d bet the house on it — when that powerpoint slide comes up, someone on your committee is going to ask about loading controls.

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Topics: Protocols, Antibody Performance, Western Blot, Companion Reagents, techniques

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

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: Protocols, Antibody Performance, Primary Antibodies, IF-IC, techniques

Journal Club | Zika virus turns off Akt signaling to hijack autophagy in developing neural tissue


Posted by Ken B on Mar 29, 2017 3:00:00 AM

The Zika virus (ZIKV) is related to flaviviruses that cause dengue and yellow fever, and is spread via mosquito (Ae. aegypti and Ae. albopictus) bites and secondarily via sexual transmission. ZIKV infection in adults is associated with mild dengue-like disease or Guillain-Barré syndrome, an autoimmune disorder of the peripheral nervous system. Most alarmingly, ZIKV is also transmitted across the placental barrier during pregnancy and can lead to the severe birth defect microencephaly (underdevelopment of the brain).

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Topics: Journal Club, Literature Review

Successful Immunofluorescence: Fixation and Permeabilization


Posted by Ken B on Mar 15, 2017 3:00:00 AM

Part Three of a four-part series on Immunofluorescence. Check out The Importance of Validation and Experimental Controls.

The performance of an antibody is a crucial determinant in getting reliable immunofluorescence (IF) results. Equally important is the preparation of the biological sample - cells or tissue used in your experiments - before any antibodies are introduced. The fixation and permeabilization of your samples are key steps that can determine your experiment’s failure or success. The ideal fixative preserves a “life-like” snapshot while quickly stopping the degradative process of autolysis by crosslinking and inhibiting endogenous enzymes. This post provides examples of how different antibodies perform at their best using different protocols.

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Topics: Protocols, Antibody Performance, IF-IC, techniques, Fixation

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