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.
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.
A Message from our Chief Scientific Officer, Roberto Polakiewicz, Ph.D.
The time is now for the scientific community to come together to respond to the challenges of reproducibility and to be part of the solution . . .
CRISPR, a nifty gene-editing tool, has swept the scientific community off its feet. It has a catchy name, it’s in every other publication, it is the subject of a volatile legal dispute involving claims to its intellectual property (1), its use has instigated a multitude of summits to do with its ethical ramifications and, despite the fact that it’s less than 4 years old, the Science journal named it the ‘breakthrough of the year’ (2). To give you some perspective, the other contenders were the New Horizon’s epic visit to the dwarf planet Pluto, the discovery of a lymphatic system in the brain, and the Ebola vaccine.
Well, you get the picture, CRISPR is a big deal, but what on Earth is it and why are scientists so smitten by it?
Antibody Validation Roundtable: Specificity + Sensitivity + Reproducibility = Validation
Concerns about the use of incorrectly or insufficiently characterized antibodies in biomedical research have been raised by recent reports of nonreproducible research studies. Non-reproducible research studies have resulted in loss of time, resources, and scientific reputation, with publication retractions occurring in certain cases.