When it comes to diversity, cytokines give scientists many options to explore. These small proteins—often just 5 kDa to 20 kDa—carry signals from one cell to another, and these messages can call out the immunity troops to battle infection, inflammation and more. Cytokines also come in many forms: peptides, proteins and glycoproteins. That assortment of structures demands flexible methods of analysis, such as the ELISA.
In brief, an ELISA uses an antibody that targets a specific antigen. When the two bind, an ELISA generates some indicator, such as a change in color or a fluorescent or chemiluminescent signal, which can be detected with a spectrophotometer or other device. Typically, scientists run ELISAs in 96- or 384-well plates. As Joshua Magnuson, product manager at Bio-Techne, points out, “The ELISA format, despite being in use for over 40 years, remains a top choice to monitor and characterize cytokines for a number of reasons, starting with accurate quantitation.”
As Magnuson explains, “The ELISA’s focus on a single cytokine allows the assay developer to deal with binding proteins and ligands without concern for how it might affect other cytokines in the sample.” Moreover, it doesn’t take much equipment to run an ELISA. It can be performed with only a plate reader. In addition, this assay is easy, and it “can be performed by any level of researcher and achieve consistent, reproducible results,” Magnuson says.
As tool providers continue to optimize and develop reagents, scientist can expect to perform more detailed analysis on cytokines. Jeremy Lambert, marketing director of Quanterix also shared “Recent developments in ultra-sensitive single-molecule detection of cytokines has enabled researchers to quantitatively measure cytokines with >1000X improvement in sensitivity.” He further adds that “advancement in detection technologies have taken advantage of improved quality of antibody reagent and specifically antibodies with slower off-rates which tend to perform best with ultra-sensitive assays.”
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