Glucagon Kit
Glucagon quantification in cell supernatant
Glucagon Serum Kit HTRF®
The Glucagon kit is intended to provide significant time-saving over ELISA, in the quantification of glucagon in serum and plasma samples.
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Low sample consumption
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Faster and more convenient than ELISA
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Scalable from 96 to 1536 format
Overview
The Glucagon offers superior specificity and sensitivity for a reliable quantification of glucagon in serum and plasma samples. Built on a truly homogeneous protocol, the assay does not require any washing or complex extraction steps, meaning significant time savings over ELISA methods. Simply add the detection reagents, incubate, and read!
Benefits
- HIGH SPECIFICITY
- HIGH SENSITIVITY
Assay principle
Glucagon is measured using a sandwich immunoassay with two monoclonal antibodies, one labelled with Lumi4-Tb Cryptate (donor) and the other with d2 (acceptor). The intensity of the FRET signal obtained is proportional to the concentration of glucagon in the sample.
Assay protocol
10 µL of cell supernatants are supplemented with 5 µL of acceptor-labelled antibodies and 5 µL of donor-labelled antibodies (or 10 µL of pre-mixed reagents for a single dispensing step). The mix is then incubated from 4h to overnight at RT and results are read on an HTRF compatible reader.
Specificity
| Peptide | Specificity ( % of recognition ) |
|---|---|
| Glucagon | 100% |
| Oxyntomodulin | <0.07% |
| Glicentin | <0.07% |
| Glucagon fragment 1-18 | <1.81% |
| Glucagon fragment 19-29 | <0.03% |
| GLP-1 (7-36) amide | <0.06% |
| GLP-1 (7-37) | <0.11% |
| GLP-2 | <0.3% |
| GRPP (Glicentin-Related Pancreatic Peptide) | <0.01% |
Sensitivity
Typical calibration curves obtained by the dilution of glucagon standard in cell-culture media conditions for 20 µL final, using a white 384-well small volume plate, are presented below. Samples were incubated overnight at RT and diluted in diluent #6.
The assay ranges from 15,6 to 2000 pg/mL and the detection limit measured was 12 pg/mL (PHERAstar FS reader - flash lamp excitation).
Linearity
Dilution linearity in cell culture media was assessed, with recovery results between 100% and 117%.
Human serum samples were collected and serially diluted in diluent #6. The results are expressed as percent observed from expected.
Serial dilutions of a human serum sample in diluent #6: Human serum sample [Glucagon] measured (pg/mL) [Glucagon] expected (pg/mL) % of expected concentration Undiluted 1666.7 - - 1:2 846.3 833.3 101.6 % 1:4 420.7 416.7 101.0 % 1:8 221.7 208.3 106.4 % 1:16 121.8 104.2 116.9 %
Intra- and inter-assay variability
Intra-assay and inter-assay variability in cell culture media conditions were assessed. Typical CVs of less than 4% for intra- and less than or around 10% for inter- were obtained, supporting claims regarding the features of a robust and reproducible assay.
Intra-Assay - n = 24 | CV |
|---|---|
Standard 2 (= 31.25 pg/mL) | 1.8% |
Standard 6 (= 500 pg/mL) | 1.9% |
Standard 8 (= 2,000 pg/mL) | 3.6% |
Intra-Assay - n = 6 | CV |
|---|---|
Standard 2 (= 31.25 pg/mL) | 11.2% |
Standard 6 (= 500 pg/mL) | 6.6% |
Standard 8 (= 2,000 pg/mL) | 6.4% |
Normal and pathological levels of glucagon
Glucagon concentrations for 16 pathological and normal EDTA-plasma samples were determined using the HTRF Serum Glucagon kit reagents. The results obtained show the good discrimination of the glucagon assay, clearly differentiating between normal and pathological levels of glucagon.
Product Insert Glucagon (serum) Kit / 62SGLPEB-62SGLPEF
62SGLPEB-62SGLPEF - Product Insert
HTRF Product Catalog 2020 July update
All your HTRF assays in one document! - Catalog
A guide to Homogeneous Time Resolved Fluorescence
General principles of HTRF - Guides
A guide to help you in your diabetes projects
A comprehensive listing of Type II diabetes pathways - Guides
How HTRF compares to Western Blot and ELISA
Get the brochure about technology comparison. - Brochures
Safety Data Sheet (DEU) Glucagon (serum) Kit / 62SGLPEB
62SGLPEB - Safety Data Sheet
Safety Data Sheet (FRA-FR) Glucagon (serum) Kit / 62SGLPEB
62SGLPEB - Safety Data Sheet
Safety Data Sheet (ITA) Glucagon (serum) Kit / 62SGLPEB
62SGLPEB - Safety Data Sheet
Safety Data Sheet (SPA) Glucagon (serum) Kit / 62SGLPEB
62SGLPEB - Safety Data Sheet
Safety Data Sheet (ENG-GB) Glucagon (serum) Kit / 62SGLPEB
62SGLPEB - Safety Data Sheet
Safety Data Sheet (ENG-US) Glucagon (serum) Kit / 62SGLPEB
62SGLPEB - Safety Data Sheet
Safety Data Sheet (DEU) Glucagon (serum) Kit / 62SGLPEF
62SGLPEF - Safety Data Sheet
Safety Data Sheet (ELL) Glucagon (serum) Kit / 62SGLPEF
62SGLPEF - Safety Data Sheet
Safety Data Sheet (FRA-FR) Glucagon (serum) Kit / 62SGLPEF
62SGLPEF - Safety Data Sheet
Safety Data Sheet (ITA) Glucagon (serum) Kit / 62SGLPEF
62SGLPEF - Safety Data Sheet
Safety Data Sheet (SPA) Glucagon (serum) Kit / 62SGLPEF
62SGLPEF - Safety Data Sheet
Safety Data Sheet (ENG-GB) Glucagon (serum) Kit / 62SGLPEF
62SGLPEF - Safety Data Sheet
Safety Data Sheet (ENG-US) Glucagon (serum) Kit / 62SGLPEF
62SGLPEF - Safety Data Sheet
Plate Reader Requirement
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