A high quality enzyme immunoassay for sensitive and specific determination of glucagon in rat, mouse, and non-human primate (NHP) samples.
Read the Directions for Use
10 μL sample volume
No purification of samples
Little or no cross-reactivity to peptides oxyntomodulin, glicentin, mini-glucagon, GLP-1, GLP2 and GRPP.
Glucagon White papers:
Mercodia Glucagon ELISA - 10 μL provides a method for the quantitative determination of glucagon in rat, mouse, and non-human primate (NHP) serum, EDTA plasma and cell culture media samples.
Format: 1 x 96 wells
Specificity: Complete table of cross reactivity
Samples: Serum, EDTA plasma and cell culture medium samples
Sample volume: 10 μL
Assay range: 2-180 pmol/L
Detection limit: ≤ 1.5 pmol/L
Incubation: 18-22 h (overnight) + 15
The detection limit is ≤ 1.5 pmol/L (5.2 pg/mL)
as determined with the methodology described in ISO11843- Part 4.
Recovery and Precision
see the product sheet
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.
For studies in which very low levels of glucagon need to be detected, it may be beneficial to use BD (Becton Dickinson) P800 tubes for sample collection, since this will prevent the degradation of glucagon. If possible, avoid storing samples at room temperature or 2-8°C for long periods of time.
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.
Summary of protocol
|Glicentin, human||1.0 %|
|Glicentin, mouse||7.0 %|
|Glicentin, rat||4.0 %|
|Oxyntomodulin, human/rat/mouse||2.0 %|
n.d.= not detected