Protocols and Tips

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.

We asked core facilities staff and research scientists from the U.S. and Europe for their top tips for securing funding for large instrumentation, such as a cell analyzer or cell sorter. Here are the key questions you should consider while preparing your grant application to increase your chances of a successful outcome.

Mice are critical model organisms for the study of neuroinflammation. But tissue quantity is often a limiting factor. In a recent paper, researchers describe a protocol for analysis of 33 cytokines and chemokines using only 50 μl of tissue homogenate. The remaining homogenate can be used for RNA extraction or other analysis.

Presented here are some tips to help you avoid buyer’s remorse and make an educated decision about which instrument to purchase. These tips provide you with the tools to make an informed decision when purchasing a cell sorter.

Incorporating fluorescence-based cell sorting into your lab grants you powerful access to new research areas and rarer cell types without sacrificing efficiency or results. New, powerful techniques like CRISPR and single-cell analysis are enhanced by the isolation of specific cells. But isolating cell populations of the right quality and quantity is both challenging and time consuming. We offer some tips to set yourself up for success and get the most out of your sorts.

Protein thermal shift assays enable quick and easy buffer optimization for increased protein stability. See how Bio-Rad’s family of CFX Real-Time PCR Detection Systems can measure protein thermal stability with higher throughput and more buffer systems than traditional circular dichroism detection.

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.

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.