DA-E 5090

By lysing cells, releasing RNA, and removing impurities such as proteins and DNA, high-purity RNA products are finally obtained. The commonly used traditional method is the guanidine isothiocyanate/phenol/chloroform method (Trizol), which is suitable for a variety of animal materials including animal tissues, microorganisms, cultured cells, etc., and most plant materials.

Enzyme-linked immunosorbent assay (ELISA) is a biochemical technology widely used to detect and quantify proteins, antibodies, hormones and other biomolecules. This method uses enzyme-labeled antibodies or antigens to specifically bind to the test substance, and generates a measurable signal through an enzyme-catalyzed reaction. ELISA has the characteristics of high sensitivity, specificity and high-throughput, and is suitable for medical diagnosis, biomedical research and drug development and other fields.

Northern blot detects a defined RNA species by separating denatured RNA by size, transferring RNA to a membrane, hybridizing with a complementary labeled DNA or RNA probe, and detecting probe-bound RNA by autoradiography, phosphorimaging, or validated nonradioactive detection. The readout is both RNA size and abundance: band migration estimates transcript length or RNA-processing state, while band intensity reflects relative target RNA amount after normalization to total RNA, rRNA, or another validated loading control. In cancer cells, primary neurons, mouse tumor samples, intestinal organoids, inflammatory macrophages, or drug-screening studies, Northern blot is most appropriate when transcript size, isoform pattern, RNA processing, or small-RNA detection is important; qPCR or RNA-seq can complement it when higher sensitivity or global profiling is needed.

Pyroptosis is a lytic inflammatory cell-death pathway executed by gasdermin pores, most classically through inflammasome-mediated activation of caspase-1, cleavage of gasdermin D, membrane pore formation, LDH release, and secretion of IL-1β and IL-18. The canonical pathway is commonly modeled by priming cells with an inflammatory signal such as LPS to induce pro-IL-1β and inflammasome components, followed by an activation signal such as ATP or nigericin to activate NLRP3, ASC speck formation, caspase-1 cleavage, GSDMD cleavage, cytokine release, and pyroptotic membrane rupture. The non-canonical pathway is triggered when cytosolic LPS activates mouse caspase-11 or human caspase-4/5, leading to GSDMD cleavage and pyroptosis, and this can secondarily activate NLRP3-dependent IL-1β release. Pyroptosis is linked to inflammatory injury, infection, cancer, liver disease, ocular disease, placental inflammation, and other disease phenotypes, but unresolved questions include which gasdermin fam