When you’re shopping for antibodies, there are so many factors to consider. For example, will it work in my cell or tissue model? Has it been tested in the application I want to use? Sometimes it’s a struggle to find what you need because your options are limited, but in other instances there may be several reagents that seem like they could work in your experiment.
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.
I’m a Product Scientist at Cell Signaling Technology (CST). My typical work day no longer looks like my fellow scientists’ daily grind, and at times, I even find it difficult to describe in words what I do, but I’ll give it a shot!
Here at CST, we frequently get questions about the proper shipping temperature for antibodies. So we thought we would do an interview with an expert on this topic, our very own Seneca Stone who directs our global supply chain operations.
We as scientists learn from each success and failure. Sometimes it takes many failures to achieve success. And some discoveries are made with no fanfare, far from the spotlight. Other times, a good day’s work is even sweeter when you realize someone noticed!
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...”
The importance of antibodies as tools in scientific research studies cannot be understated, yet these reagents have increasingly come under fire for their lack of reproducibility. Part of the issue is that the antibody market is composed of hundreds of vendors and resellers with varying definitions for validation and consistency. Cell Signaling Technology (CST) believes that antibody suppliers should be held accountable for the products they provide, but that vendors alone cannot solve the reproducibility “crisis." How antibodies are validated and used in the laboratory is a critical component to this process. Researchers need to be more attentive to following established protocols and leverage the expertise of the scientists who have developed and tested the product they are using. Journals need to be more active in enforcing existing policies regarding materials and methods or develop more clear-cut means to identify and describe the use of biological reagents in published research. During this webinar we will address the role vendors, researchers, and journals should play in minimizing irreproducibility. We will also outline CST’s antibody validation process, while highlighting steps all users should consider when selecting and using antibodies in their research.
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.
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.
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.