A high quality enzyme immunoassay for sensitive and specific determination of glucagon in rat, mouse, porcine and non-human primate (NHP) samples. Read the Directions for Use
Read our White Paper on the challenges of measuring glucagon.
Mercodia Glucagon ELISA - 10 μL provides a method for the quantitative determination of glucagon in rat, mouse, porcine and non-human primate (NHP) serum, EDTA plasma and cell culture media samples.
Format: 1 x 96 wells
Samples: Serum, EDTA plasma and cell culture medium
Sample volume: 10 μL
Assay range: 2-180 pmol/L
Detection Limit: 1.5 pmol/L
Incubation: 18-22 h (overnight)
Keep samples cold during analysis. Serum, EDTA plasma and cell culture medium can be used. However, glucagon in serum or EDTA plasma samples will be sensitive to storage conditions and freeze-thaw cycles. Addition of aprotinin to EDTA plasma samples will not improve stability.
Collecting samples in Becton Dickinson (BD) P800 tubes containing lyophilized protease inhibi- tors and DPP-IV inhibitors will yield (20 – 30%) higher glucagon values than EDTA plasma not collected in BD P800 tubes or serum because of improved stability. Samples from P800 tubes are stable for up to 6 hours at room temperature or 2-8°C, and up to 4 freeze-thaw cycles in cryo vials. Store samples at -80°C.
Grossly lipemic or icteric samples do not interfere in the assay. Samples with high levels of hemoglobin (>500 mg/dl) can interfere in the assay.
Mercodia Glucagon ELISA – 10 μL is a solid phase two-site enzyme immunoassay. It is based on the direct sandwich technique in which two monoclonal antibodies are directed against separate antigenic determinants on the glucagon molecule. During incubation glucagon in the sample reacts with peroxidase-conjugated anti-glucagon antibodies (clone E6A11K) and anti-glucagon antibodies (clone M5F9S) bound to microplate wells. A simple washing step removes unbound enzyme labelled antibody.The bound conjugate is detected by reaction with 3,3’,5,5’-tetramethyl-benzidine (TMB). The reaction is stopped by adding acid to give a colorimetric endpoint that is read spectrophotometrically.
Tested conc. (pmol/L)
|Oxyntomodulin. bovine, canine, porcine||n.d.||400|
1. Wewer Albrechtsen, N, J, Hartmann, B et al., (2014)
Hyperglucagonaemia analysed by glucagon sandwich ELISA: nonspecific interference or truly elevated levels?
2. Chow, S, Z, Speck, M et.al., (2014)
Gp130 receptor signaling mediates alpha cell dysfunction in a rodent model of type 2 diabetes
3. Malmgren, S, Ahren, B, (2015)
DPP-4 inhibition contributes to the prevention of hypoglycaemia through a GIP-glucagon counterregulatory axis in mice