Human GLP-2 ELISA Assay
The Human GLP-2 ELISA Assay is For Research Use Only
Size: 1×96 wells
Dynamic Range: 0.412 – 100 ng/mL
Incubation Time: 20 hours
Sample Type: Serum, Plasma
Sample Size: 25 µL
Specificity: This Human GLP-2 ELISA is highly specific to human GLP-2 and shows cross-reactivity to neither glucagon (rat/mouse/human) nor GLP-1 even at a concentration of 300 pmol/mL.
Alternative Name: Glucagon-like Peptide 2
Stability and Storage
Storage: Store all the components at 2~8°C.
Shelf: life The kit is stable under the condition for 19 months from the date of manufacturing. The expiry date is stated on the package.
Package: For 96 tests per one kit including standards.
This Human GLP-2 ELISA Assay kit for determination of human GLP-2 in plasma and serum samples is based on a competitive enzyme immunoassay using combination of highly specific antibody to human GLP-2 and biotin-avidin affinity system. To the wells of the plate coated with goat anti rabbit IgG, standard antigen or samples, biotinylated human GLP-2, and rabbit anti GLP-2 antibody are added for competitive immunoreaction. After incubation and plate washing, horse radish peroxidase (HRP) labeled streptoavidin (SA) is added to form HRP labeled SA – biotinylated GLP-2 – antibody complex on the surface of the wells. Finally, HRP enzyme activity is determined by o-phenylenediamine dihydrochloride (OPD) and the concentration of human GLP-2 is calculated.
Rat GLP-2 ELISA Assay
Mouse GLP-2 ELISA Assay Kit
Human GIP (Active) ELISA Assay Kit
Proglucagon gene is expressed in both pancreatic A cell and intestinal L cell. Tissue-specific posttranslational processing of proglucagon by prohormone convertase produces different proglucagon derived peptides (PGDPs) in both pancreas and intestine. The most notable pancreatic PGDP is glucagon, whereas intestinal L cell produces several structurally related peptides, including glucagon-like peptide 1 (GLP-1) and 2 (GLP-2), as well as glicentin and oxyntomodulin which contain glucagon sequence in their molecules. Among PGDPs, GLP-2 has been found to show intestinal epithelial proliferation.
- Bring all the reagents and samples to room temperature (20-30°C) at least 1 hour before starting assay.
- Add 0.35mL/well of washing solution into the wells of the plate, and then aspirate the solution. Repeat this washing procedure further twice (total 3 times). Finally, invert the plate and tap it onto an absorbent surface, such as paper toweling, to ensure blotting free of most residual washing solution.
- Fill 40uL of labeled antigen solution into the wells first, then introduce 25uL each of standard solutions (0, 0.412, 1.235, 3.704, 11.11, 33,33 and 100 ng/mL) or samples and finally add 50uL of GLP-2 antibody into the wells.
- Cover the plate with adhesive foil and incubate it at 4ºC for 16 ~ 18 hours. (Not shaken)
- After incubation, take off the adhesive foil, aspirate the solution in the wells and wash the wells 3 times with approximately 0.35 mL/well each of washing solution. Finally, invert the plate and tap it onto an absorbent surface, such as paper toweling, to ensure blotting free of most residual washing solution.
- Pipette 100uL of SA-HRP solution into each of the wells.
- Cover the plate with adhesive foil and incubate it at room temperature (20~30ºC) for 1 hour. During incubation, the plate should be shaken with a microtiter plate shaker.
- Resolve one OPD tablet with 12 mL of substrate buffer. It should be prepared immediately before use.
- Take off the adhesive foil, aspirate the solution in the wells and wash the wells 5 times with approximately 0.35 mL/well each of washing solution. Finally, invert the plate and tap it onto an absorbent surface, such as paper toweling, to ensure blotting free of most residual washing solution.
- Add 100uL of substrate solution into each the wells, cover the plate with adhesive foil and incubate it for 30 minutes at room temperature.
- Add 100uL of stopping solution into each of the wells to stop color reaction.
- Read optical absorbance of the solution in the wells at 490 nm. Calculate mean absorbance values of standard solutions and plot a standard curve on semi logarithmic graph paper (abscissa: concentration of standard solution; ordinate: absorbance value). Use the average absorbance of each sample to determine the corresponding value by simple interpolation from this standard curve.
Typical Standard Curve
Precision and Reproducibility: