You've just finished running your lysate sample by SDS-PAGE, so you're pretty confident your protein should have migrated at the right molecular weight and is ready to transfer to a membrane for detection by Western Blot. But what if it's not?
The preparation of a cell lysate is crucial to the success of many assays, including techniques that use antibodies to characterize protein expression. Ideally, throughout the whole process of sample preparation, from lysate collection to analysis, your protein of interest would be well-preserved and protected from the activities of proteases and phosphatases. Proper storage and handling of the lysate are crucial to avoiding degradation and maintaining the ability to detect your protein.
Western blotting is used to detect proteins and analyze their expression using antibodies, but it does have its limitations. Flow cytometry also employs antibodies, but it uses fluorescence to detect and analyze protein expression on a cell-by-cell basis.
The success of your chromatin immunoprecipitation (ChIP) experiment depends on the fragmentation of chromatin, a critical step in the ChIP protocol. This can be accomplished with either sonication or enzymatic digestion. But how do you decide which chromatin fragmentation protocol to use in your ChIP experiments? A number of factors can influence your choice, making a decision seem daunting. So let’s simplify – watch the video and we'll will show you the way!
Understanding the microenvironment of disease is critical. Targeted cancer therapies may not be effective depending on the combination of cell types in the tumor. Likewise, an immune cell invasion in pancreatic tissue could indicate the progression of type 1 diabetes. For years, this complex landscape could only be witnessed in single-cell snapshots by flow cytometry or limited tissue staining. Imaging Mass Cytometry™ (IMC™) now offers a panorama of this dynamic world.
Culturing cells in the lab? Following these tips will add to your success and help you avoid wasted time or the dreaded cell contamination!
Cell lines are a crucial part of life science research and development. But did you know an estimated 18-36% of cell lines are believed to be misidentified or cross-contaminated with another cell line?
Running an ELISA can be a pain. Identifying pairs for an ELISA is a tedious business, and that’s before developing and validating the ELISA assay itself. Using a kit can simplify the process, but at what cost? Will that kit hinder reproducibility by introducing lot-to-lot variability over the course of my project’s lifetime? Many kits still require numerous reagent addition, incubation, and wash steps that add hands-on time and complexity to your assay.
Flow cytometry enables quantitative analysis of protein expression, signaling states, and physical characteristics (cell size/granularity) at the single-cell level. Modern flow cytometers are capable of collecting data on multiple proteins from thousands of cells per second in a heterogeneous mixture. While flow cytometry is commonly employed to identify cell types using phenotypic markers expressed on the cell surface, it can also be used to measure intracellular signaling events.