Multiple analytical obstacles have made accurate and reliable measurement of glucagon difficult. In the white paper Glucagon Measurement - Addressing Long-Standing Analytical Challenges these challenges are discussed, publications detailing researchers’ needs are reviewed and two novel glucagon assays are introduced, providing analytical solutions for the research community.
The second white paper, Glucagon - Metabolic Regulator Critical for Pathophysiology and Pharmacology, focuses on the role of glucagon and why it should be considered as a metabolic marker.
Sample volume requirements have significantly limited the use of glucagon assays. Most commercially available methods require at least 50-100uL of plasma. This has direct implications on the number of analytes that can be measured, the number of time points that can be examined and thus, the scope of scientists’ conclusions. Mercodia offers 2 very low sample volume assays and assay choice depends on sample type.
Human Samples: Mercodia Glucagon ELISA (10-1271-01)
Animal Samples: Mercodia Glucagon ELISA - 10uL (10-1281-01)
In a subsequent study, the new Mercodia assay (10-1271-01) was included and based on the authors’ extensive evaluation and expertise in this field, they determined that “the Mercodia assay has the best performance, in terms of specificity, precision and sensitivity data”. They concluded that only the Mercodia assay was “suitable for measuring glucagon concentrations in clinical samples”. Wewer Albrechtsen NJ et al. (2014) Diabetologia 57:1919-1926
BEST SENSITIVITY ON THE MARKET
Even though the physiological range of glucagon is reported to be 0 – 30 pmol/L (0 – 105 pg/mL), for many years, commercially available glucagon assays did not provide sufficient sensitivity for detecting glucagon under a variety of physiological conditions. Experts in the field state: “Clearly, assays with sensitivities >5pmol/L are, therefore, unsuitable for the complete characterization of glucagon secretion.” The sensitivities of Mercodia’s two assays, coupled with broad dynamic ranges, afford scientsts the ability to measure physiologically relevant concentrations of glucagon in a variety of experimental paradigms.
GLUCAGON-SPECIFIC MONOCLONAL ANTIBODIES
While glucagon’s actions may be unique, its sequence is not (see above). The use of an assay that cross-reacts with other glucagon-containing proteins may result in inaccurate characterization of alpha cell function and glucagon kinetics under various physiological conditions, as well as misinterpretation of how a subject is responding to therapy.
Mercodia’s glucagon assays are based on 2 highly specific monoclonal antibodies that recognize the terminal ends of glucagon.
ASSAYS FOR TRANSLATIONAL RESEARCH
Glucagon’s sequence is highly conserved across species, but validation of different sample types is crucial because sequence homology is not the only factor that affects antibody-antigen binding in an immunoassay (e.g., matrix interferences). Mercodia’s assays contain a unique blocking solution to prevent or minimize matrix interferences.
In addition, both of the Mercodia glucagon assays are based on the same 2 monoclonal antibodies, ensuring the same specificity for translational sample analyses. Mercodia’s glucagon assays can be used with cell culture, rodent, canine, porcine, nonhuman primate and human samples.
(See the Low Sample Volume section above for information on choosing the right Mercodia glucagon assay.)
Glucagon is essential to glycemic control but also has many other important biological connections...