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.

READ MORE >

Topics: Protocols, Western Blot, techniques, Tech Tips

Mastering Phospho-Flow: Immune Cell Activation Webinar


Posted by Chris S on Jul 11, 2018 3:15:00 AM

To characterize activation of immune cells, measurement of intracellular signaling using phospho-specific antibodies combined with immune phenotyping using surface markers is ideal. Owing to the unique capability of flow cytometry to measure events at a single-cell level, “Phospho Flow” allows for the identification of rare signaling events during processes such as co-stimulatory signaling and manipulation of checkpoint inhibition. However, combining extracellular and intracellular antibodies in a flow protocol can present technical challenges. 
 
READ MORE >

Topics: Flow, Protocols

Are you ready to transfer your protein? Wet vs. semi-dry transfer


Posted by Neha G on Apr 18, 2018 3:15:00 AM

 You are all amped up to run a western blot to identify “your favorite protein.” The lysates have been run and proteins separated by SDS-PAGE. Now it’s time to transfer proteins from the gel to the membrane, and you’re sitting wondering….wet or semi-dry?? Or maybe you are better prepared than I was as a graduate student and you already know your next step, in which case you are aware of the pros and cons of wet and semi-dry transfer.

READ MORE >

Topics: Science Education, techniques, Western Blot, Protocols

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: ChIP, Antibody Performance, Protocols, Reproducility, techniques

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.

READ MORE >

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

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.

READ MORE >

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

5 Steps to Publication-Worthy IF Images


Posted by Tara W on Sep 6, 2017 3:30:00 PM

A picture is worth a thousand words, or in the case of immunofluorescent imaging, a thousand proteins. The images used to illustrate a scientific experiment should convey as much information as the text itself. Here at CST, we pride ourselves in the quality of our antibodies and our rigorous validation process. When we approve our primary antibodies for IF, we like to showcase them using high quality images generated in-house. Beyond our recommended IF protocols (check it out here), here are some additional considerations to make when planning your IF staining. 

READ MORE >

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

Switching from Manual to Automated IHC? We've got your back


Posted by Chris G on Aug 30, 2017 3:00:00 AM

If you’ve ever transitioned your IHC experiments from a manual protocol to an automated platform, you may have found the conversion process to be a drag. It isn’t an easy thing to do. For that reason, we’re happy to announce our IHC Leadership in Automation initiative. This rigorous validation initiative expands on our already thorough measures, allowing researchers to not only use CST products with our recommended manual IHC protocol, but also to bridge the assay to new platforms and techniques. Our foray into the world of automated IHC aims to reduce the amount of time researchers spend on assay transfer and protocol optimization.

READ MORE >

Topics: Antibody Performance, Antibody Validation, IHC, Automated IHC, Protocols, techniques

Intracellular Flow Cytometry in Action


Posted by Liana G on Aug 9, 2017 9:45:00 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.

READ MORE >

Topics: Protocols, Flow, Cell Biology, techniques

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


Posted by Carolyn P on May 31, 2017 3:00:00 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.

READ MORE >

Topics: Protocols, IHC, techniques

Subscribe to Email Updates

Recent Posts