Mouse GLP-2 ELISA Assay kit:
For Research Use Only
Size: 1×96 wells
Dynamic Range: 0.412 – 100 ng/mL
Incubation Time: 20 hours
Sample Type: Mouse serum, plasma
Sample Size: 25μL
This Eagle Biosciences Mouse GLP-2 ELISA Assay kit is used for quantitative determination of mouse GLP-2 in serum or plasma samples. The kit is characterized for sensitive quantification, high specificity and no influence with other components in serum or plasma and needlessness of sample pre-treatment. Mouse GLP-2 standard is highly purified synthetic product. The Eagle Biosciences Mouse GLP-2 ELISA Assay is for Research Use Only and should not be used for diagnostic procedures.
The proglucagon gene is expressed in both pancreatic A cell and intestinal L cell. Tissue-specific posttranslational processing of proglucagon by the prohormone convertase produced the different proglucagon derived peptides (PGDPs) in both pancreas and intestine. The most notable pancreatic PGDP is glucagon, whereas the L cell produces several structurally related peptides, including glucagon-like peptide(GLP)-1 and GLP-2, as well as glicentin and oxyntomodulin, which contain glucagon sequence in their molecules. Among PGDPs, GLP-2 has recently been found to show intestinal epithelial proliferation.
This Eagle Biosciences Mouse GLP-2 ELISA Assay Kit for determination of mouse GLP-2 in serum or plasma samples is based on a competitive enzyme immunoassay using combination of highly specific antibody to rat GLP-2 (strong cross reactivity to mouse GLP-2) and biotin-avidin affinity system. The 96-wells plate is coated with goat anti rabbit IgG antibody. Mouse GLP-2 standard or samples, labeled antigen and anti-rat GLP-2 polyclonal antibody are added to the wells for competitive immunoreaction. After incubation and plate washing, HRP labeled streptoavidin (SA-HRP) are added to form HRP labeled streptoavidin-biotinylated rat 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 mouse GLP-2 is calculated.
- Bring all the reagents and samples to room temperature before beginning the test.
- Fill 75uL of labeled antigen solution into the wells first, then introduce 25uL of each of standard solutions (0, 0.412, 1.235, 3.704, 11.11, 33,33, 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.(Still, shaker not need)
- Take off the adhesive foil, aspirate the solution in the wells and wash the wells three times with approximately 0.35 mL/well of washing solution.
- Pipette 100uL of SA-HRP solution into the wells.
- Cover the plate with adhesive foil and incubate it at room temperature (20-30ºC) for 1 hour. During the incubation, the plate should be shake with a plate shaker.
- Resolve OPD tablet with 12 mL of substrate buffer. It should be prepared immediately before use.
- Take off the adhesive foil, aspirate and wash the wells five times with approximately 0.3 mL/well of washing solution.
- Add 100uL of substrate solution into the wells, cover the plate with adhesive foil and incubate it for 30 minutes at room temperature.
- Add 100uL of stopping solution into the wells to stop reaction.
- Read the optical absorbance of the wells at 492nm. Calculate mean absorbance values of wells containing standards and plot a standard curve on semilogarithmic graph paper (abscissa: concentration of standard; ordinate: absorbance values). Use the standard curve to read mouse GLP-2 concentrations in samples from the corresponding absorbance values.
Typical Standard Curve
This Eagle Biosciences Mouse GLP-2 ELISA Assay kit has high specificity to mouse GLP-2 and shows no cross reactivity with mouse glucagon and mouse GLP-1 even in the concentration of 300 pmol/mL.
- Philippe J.: Structure and pancreatic expression of the insulin and glucagon genes. Endocr Rev 12: 252-271,1991
- Mojsov S. et al: Preproglucagon gene expression in pancreas and intestine diversifies the level of post-transcriptional processing. J Biol Chem 261: 11880-11889,1986
- Drucker D.J. et al: Induction of intestinal epithelial proliferation by glucagon-like peptide 2. Proc Natl Acad Sci USA 93: 7911-7916,1996
- Brubaker PL, et al: Circulating and tissue forms of intestinal growth factor, glucagon-like peptide 2. Endocrinology 15: 673-677,1997
- Drucker D. J.: Perspective in diabetes Glucagon-like peptides. Diabetes 47: 159-169, 1998
- Xiao Q, et al: Secretion of the intestinotropic hormone glucagon-like peptide 2 is differentially regulated by nutrients in humans. Gastroenterology 117: 99-105,1999
- Kato I, et al: Synthesis of Rat Glucagon-like peptide(GLP)-2 and its biological and immunochemical studies. Peptide Science 1999: N.Fujii(Ed). The Japanese Peptide Society (2000)