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Immunoassays | Metabolism

Erennaź Total GLP-1(MP-based) Immunoassay Kit (Cat# 03-0025-xx)

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The Erennaź Total GLP-1 Immunoassay Kit uses a quantitative fluorescent sandwich immunoassay technique to measure Total GLP-1 plasma and serum samples. A capture antibody specific for the C-terminus of 7-36 (amide) and 9-36 (amide) GLP-1 has been pre-coated onto paramagnetic microparticles (MP). The user pipettes MP, standards and samples into uncoated microplate wells. During incubation, any GLP-1 present in the sample binds to the capture antibody on the coated MP. Unbound molecules are washed away during the subsequent buffer exchange and wash steps. Fluor-labeled detection antibody is added to each well and incubated. This detection antibody recognizes and binds to the N-terminus of the Total GLP-1 that has been captured onto the MP. During the following wash step the MP’s are transferred to a clean plate. Elution buffer is then added and incubated. The elution buffer dissociates the bound protein sandwich from the MP surface, releasing the labeled antibodies. These antibodies are separated during transfer to a final Microplate. The plate is loaded into the Erenna System where the labeled molecules are detected and counted. The number of fluor-labeled detection antibodies counted is directly proportional to the amount of Total GLP-1 present in the sample when captured. The amount of Total GLP-1 in unknown samples is interpolated from a standard curve.

Assay Performance

TABLE 1. Analytical sensitivity of the Erennaź Total GLP-1(MP-based) Immunoassay Kit1
Lower Limit of Detection 0.01 pg/mL
Lower Limit of Quantification2 0.4 pg/mL
Upper Limit of Quantification 200 pg/mL
Low-end CV% Range 5 - 12%
Low-end CV% Average 9%
Recommended Sample Volume 20 μL
Minimum Sample Volume Required3 5 μL

1 See product insert for updated values
2 LLoQ = 20% CV and ± 20% recovery
3 based upon median [GLP-1, total (MP-based)] in a healthy reference

FIGURE 1. [Total GLP-1] in EDTA plasma from 10 healthy donors, with median and interquartile range. The Erennaź Total GLP-1 Immunoassay Kit reliably quantifies Total GLP-1 in healthy subjects, who have a median [Total GLP-1] of 17.8 pg/mL that is well above the detection limit of 0.01 pg/mL.

FIGURE 2. Erennaź Total GLP-1 Immunoassay Kit low-end standard curve signal (left) and curve fit (right).

Representative data shown for demonstration purposes only. Individual results may vary depending upon samples tested and protocol used.

Biology and Disease

Glucagon-Like Peptide-1 (GLP-1) is a member of the incretin family of hormones, which are released by the gut into the bloodstream in response to food. Post-translational cleavage of the peptide produces active GLP-1, which is then rapidly degraded by protease dipeptidyl peptidase-4 (DPP-4). When blood glucose level is high, active GLP-1 enhances insulin secretion and also suppresses the release of glucagon. In addition, GLP-1 stimulates the brain to trigger feelings of satiety and regulates the rate of gastric emptying. Compared to healthy controls, some patients with type-2 diabetes show a modest decrease in GLP-1 release. Therapeutic GLP-1 analogs such as exenatide, or DPP inhibitors like sitagliptin, are promising strategies for treating type-2 diabetes.

TABLE 2. Additional UniProtKB/Swiss-Prot Information*
Protein Name:
UniProtKB/Swiss-Prot ID:
Protein-protein interaction database:
Alternative Names:
Gene Names:
Synonym Gene Names:
Biological Processes:
Molecular Function:

* Source Info from www.uniprot.org.

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