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Immunoassays | Metabolism

Erenna® Active GLP-1 (Plate-based) Immunoassay Kit (Cat# 03-0065-xx)

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Introduction

The Erenna® Active GLP-1 Immunoassay uses a quantitative fluorescent sandwich immunoassay technique to measure Active GLP-1 (7-36 amide) in plasma. A capture antibody specific for the C-terminus of x-36 amide GLP-1 has been pre-coated to a 384-well microplate. The user pipettes standards and samples into microplate wells and incubates the plate. During incubation, the Active GLP-1 present in the sample binds to the solid phase capture antibody. After the first incubation, the assay plate is washed to remove unbound molecules and detection antibody is added into microplate wells. This detection antibody recognizes and binds to the N-terminus (7-36 amide) of the Active GLP-1 that has been captured onto the solid phase. After washing, elution buffer is added and incubated to dissociate the bound protein sandwiches from the plate surface. A neutralization buffer is then added to the eluted antibodies. The fluorescent antibodies are now neutralized and free-floating in the wells. The plate is loaded into the Erenna System and the fluorescent molecules are counted. The number of fluorescently labeled detection antibodies counted is directly proportional to the amount of Active GLP-1 that was present in the sample when captured. The amount of Active GLP-1 in unknown samples is interpolated off of the standard curve.

Assay Performance


TABLE 1. Analytical sensitivity of the Erenna® Active GLP-1 (Plate-based) Immunoassay Kit1
Lower Limit of Detection 0.004 pg/mL
Lower Limit of Quantification2 0.1 pg/mL
Upper Limit of Quantification 50 pg/mL
Low-end CV% Range 3 - 13%
Low-end CV% Average 9%
Recommended Sample Volume 10 μL
Minimum Sample Volume Required3 10 μL

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


FIGURE 1. [Active GLP-1] from 9 pre- and post- feeding plasma samples, together with median concentrations.

FIGURE 2. FIGURE 2: Erenna® Active GLP-1 (Plate-based) Immunoassay Kit low-end standard curve signal (left) and curve fi t (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:
Function:
Biological Processes:
Molecular Function:

* Source Info from www.uniprot.org.


Related Products

Analyte Description  Catalog #
GLP-1, total (MP-based) Erenna® Total GLP-1(MP-based) Immunoassay Kit  03-0025-xx
GLP-1, total (Plate-based) Erenna® Total GLP-1(Plate-based) Immunoassay Kit  03-0066-xx
Mouse, (Plate-based) Insulin Erenna® Mouse (Plate-based) Insulin Immunoassay Kit  03-0006-xx

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