One-step multiplex RT-qPCR is a technique used to rapidly quantify multiple targets directly from RNA in a single reaction. But proper optimization and validation is essential for its success. These five tips will have you designing primers and probes, selecting reporters, and validating and optimizing your experiments like a pro.
Follow along as a new starry-eyed graduate student learns the ins and outs of life as a research scientist, gaining valuable western blotting tips and tricks. Along for the ride are a seasoned postdoctoral fellow and traditional principal investigator.
Being able to interrogate protein phosphorylation events and aberrant phosphorylation levels is critical to our understanding of various biological and pathological processes. However, detection of phosphorylated proteins in western blots can be challenging. These ten tips will help you generate robust data and cleaner blots.
Co-immunoprecipitation (co-IP) is a great way to identify target protein complexes and interacting partners. Though the technique can be highly informative, it is also challenging to master. The six tips and protocol outlined in this article will help you achieve successful co-IP results.
Extracting proteins from tissue lysates for western blotting can be a challenge. From knowing how to process your tissue to verifying protein loading to including suitable controls, there is a lot to keep track of. The ten tips in this article can help you on your way to generating reproducible and reliable blots and do better western blotting overall.
In the first two articles in our Finding a Good Antibody series, you learned how to select the best antibodies and received tips on proper antibody validation. In this third and final article, see how you can use this information to publish meaningful data when using antibodies. And get some guidelines for antibody best practices.
Finding antibodies that work for your application is just the first step in finding a good antibody. Next is antibody validation — making sure that your antibody does what you think it is supposed to. In this second article in our Finding a Good Antibody series, learn how best to go about validating your antibodies.
Even though antibodies are central to basic research as well as drug development and diagnostics, concerns with antibody validation remain high, and finding an antibody that works well for a specific application can be quite a challenge. Use these tips to select the best antibodies for your application.
Incorporating preamplification into your qPCR experiments may seem daunting, but it is a powerful way to analyze targets from limited samples. The tips in this article will help you understand how preamplification can enhance your qPCR experiments and the considerations and implications for incorporating it into your workflow.