Sample
Serum, EDTA Plasma, and cell-culture medium
Intended Use
Glucagon ELISA kit is an assay intended to measure the pancreatic hormone glucagon in plasma and serum. Glucagon measurements are used in the diagnosis and treatment of patients with various disorders of carbohydrate metabolism, including diabetes mellitus, hypoglycemia, and hyperglycemia.
Contents of Kit
Each Glucagon ELISA kit contains a plate sealer and reagents for 96 wells, sufficient for 42 samples and one calibrator curve in duplicate. The expiry date for the complete kit is stated on the outer label. The recommended storage temperature is 2-8°C.
1.Coated Plate, 1 plate, 96 wells (8-well strips), Ready for Use
Mouse monoclonal anti-glucagon coated plate.
For unused microplate strips, reseal the bag using adhesive tape, store at 2-8°C and use within 2 months.
2.Calibrators 1, 2, 3, 4, 5, 5 vials, 1000 μL, Lyophilized
Synthetic glucagon, Color coded yellow, Concentration stated on vial label.
Add 1000 μL redistilled water per vial.
Reconstituted Calibrators are stable for 1 month at 2-8°C.
If reconstituted Calibrators are to be used for longer than 1 month, aliquote and store at -20°C. Aliquoted Calibrators are stable for at least 2 months at -20°C.
Avoid repeated freeze/thaw cycles.
3.Calibrator 0, 1 vial, 5 mL, Ready for Use
Color coded yellow
4.Assay Buffer, 1 bottle, 22 mL, Ready for use
Color coded red
5.Enzyme Conjugate 11X, 1 bottle, 2.2 mL, Preparation see below.
Mouse monoclonal anti-glucagon
6.Enzyme Conjugate Buffer, 1 bottle, 22 mL, Ready for use
Color coded blue.
7.Wash Buffer 21X, 1 bottle, 50 mL
Dilute with 1000 mL redistilled water to make wash buffer 1X solution.
Storage after dilution: 2-8°C for 2 months.
8.Substrate TMB, 1 bottle, 22 mL, Ready for Use
Colorless solution
Note! Light sensitive!
9.Stop Solution, 1 vial, 7 mL, Ready for Use
0.5 M H2SO4
Storage
Store the kit at 2°C - 8 °C.
Precision
QC samples (P800) were analyzed in 4 replicates over 6 different occasions on one kitlot and one instrument system by three laboratory technicians.
Detection Range
1.5-130 pmol/L (5-453 pg/mL)
Detection Limit
The detection limit is 0.75 pmol/L (2.61 pg/mL) as determined by the methodology described in ISO11843-Part 4.
Lower Limit of Quantification, LLOQ, is 1.5 pmol/L (5.95pg/mL)as determined according to FDA/EMA guidelines.
The Upper Limit of Quantification, ULOQ, is 130 pmol/L (453pg/mL) as determined according to FDA/EMA guidelines.
General Description
Glucagon is a 29 amino acid polypeptide processed from proglucagon (residues 33-61) in pancreatic alpha cells. In intestinal L-cells proglucagon is cleaved into glicentin, corresponding to proglucagon residues no. 1-69. Glicentin can further be processed into oxyntomodulin, corresponding to proglucagon residues no. 33-69. Moreover, a fragment of glucagon corresponding to its C-terminal part (residues no. 19-29), also designated mini-glucagon, is reported to be present in the pancreas in low amounts compared to the total glucagon content.
In general, glucagon has an effect opposite that of insulin, i.e. it raises blood glucose levels. It causes the liver to convert glycogen into glucose, which is then released into the blood stream. With longer stimulation, glucagon action in the liver results in a glucose-sparing activation of free fatty acid oxidation and production of ketones. During hypoglycaemia, glucagon secretion offers a protective feedback mechanism, defending the organism against damaging effects of glucose deficiency in the brain and nerves.
Standard Curve
Conversion factor 1 pmol/L = 3.5 pg/mL
Citations
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