The incretin hormones, glucose-dependent insulinotropic polypeptide (GIP) and glucagons-like peptide-1 (GLP-1), are a group of gastrointestinal hormones that cause an increase in the amount of insulin released from the beta cells of the islets of Langerhans after ingestion of food. The intestinal peptide GIP was first isolated from porcine upper small intestine1). The sequences of porcine2) 3), bovine4) and human GIP5) have been determined, each has 42 amino acids, and the sequences is highly conserved. The porcine and bovine peptides differ from the human at two and three site, respectively. Takeda et al. have isolated a human cDNA encoding the GIP precursor and confirming that GIP belongs to the vasoactive intestinal peptide (VIP)/Glucagon/secretin family6). GIP is a gastrointestinal peptide hormone that is released from duodenal endocrine K cells after absorption of glucose or fat7). GIP is a potent releaser of insulin in experimental animals8) and in man 9,10) provided that the blood glucose is above basal level. Plasma level of GIP is elevated after an oral glucose load or a meal in normal man. This increase after a meal is below normal in newly diagnosed insulin–dependent diabetics11). It is now being recognized that GIP receptor is also expressed in organs and cells such as duodenum, small intestine, pancreatic alpha-cell, adipocyte and osteoblast. These results demonstrate GIP may have a lot of physiological effect in addition to their glucoregulatory effects12,13,14,15). GIP is rapidly inactivated by the enzyme dipeptidyl peptidase- 4 (DPP- 4) to GIP (3-42) with a blood half-life of only several minutes. DPP- 4 inhibitor can prolong the half-life of GIP, that expecting treatment of incretin effect.
The kit can be used for measurement of total GIP [both GIP (1-42) and GIP (3-42)] in mouse plasma with high sensitivity. It will be a specifically useful tool for incretin research.
This ELISA kit is used for quantitative determination of mouse total GIP in plasma and culture medium supernatant. The kit is characterized by its sensitive quantification and high specificity. In addition, it has no influence by other components in samples. GIP standard is highly purified synthetic product.
1) Antibody coated plate
3) HRP labeled antibody solution
4) Enzyme substrate solution (TMB)
5) Stopping solution
6) Buffer solution
7) Washing solution(concentrated)
8) Adhesive foil
This ELISA kit for determination of mouse total GIP is based on a sandwich enzyme immunoassay. To the wells of plate coated with highly purified mouse monoclonal antibody against mouse GIP, standards or samples are added for the 1st step immunoreaction. After the 1st step incubation and plate washing, HRP labeled antibody solution against mouse GIP is added as the 2nd step to form antibody - antigen - labeled antibody complex on the surface of the wells. After the 2nd step incubation and rinsing out excess labeled antibody, Finally, HRP enzyme activity is determined by 3,3’,5,5’-Tetramethylbenzidine (TMB) and the concentration of mouse total GIP is calculated.
Mouse GIP (Total) ELISA
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2. J?rnvall H, Carlquist M, Kwauk S, Otte SC, McIntosh CH, Brown JC, Mutt V. (1981) Amino acid
sequence and heterogeneity of gastric inhibitory polypeptide (GIP). FEBS Lett. 123, 205-210.
3. Moody,A.J., Damm Jorgensen, K.and Thim, L.(1981)Diabetologia 21, 306, abstr.
4. Carlquist M, Maletti M, J?rnvall H, Mutt V. (1984) A novel form of gastric inhibitory polypeptide
(GIP) isolated from bovine intestine using a radioreceptor assay. Fragmentation with staphylococcal protease results in GIP1-3 and GIP4-42, fragmentation with enterokinase in GIP1-16 and GIP17-42. Eur.J. Biochem. 145, 573-577
5. Moody, A. J., Thim, L. & Valverde, I. (1984) The isolation and sequencing of human gastric
inhibitory peptide(GIP). FEBS Lett. 172, 142-148
6. Takeda J, Seino Y, Tanaka K, Fukumoto H, Kayano T, Takahashi H, Mitani T, Kurono M, Suzuki T, Tobe T, et al.(1987) Sequence of an intestinal cDNA encoding human gastric inhibitory
polypeptide precursor. Proc Natl Acad Sci U S A. 84(20):7005-8.
7. Pederson, R.A. (1994) in Gut Peptides: Biochemistry and Physiology, eds, Walsh, J.H.& Dockray, G.J. (Raven, New York), pp,217-260
8. Rabinovitch, A. and Dupre, J (1974)Effect of the gastric inhibitory polypeptide present in impure pancreozymin-cholecystokinin of plasma insulin and glucagons in the rat. Endocrinology 94, 1139-1144
9. Dupre,J., Ross, S.A.,Watson, D. and Brown, J.C. (1973) Stimulation of insulin secretion by gastric inhibitory polypeptide in man. J. Clin. Endocrinol. Metab. 37, 826-828
10. Elahi, D., Andersen, D.K., Brown, J.C.,Debas, H.T., Hershcopf, R.J., Raizes, G.S., Tobin, J.D.and Andres, R.(1979) Pancreatic alpha-and-beta-cell response to GIP infusion in normal man. Am.J.Physiol. 237, E185-E191
11. Krarup T, Madsbad S, Moody AJ, Regeur L, Faber OK, Holst JJ, Sestoft L.(1983) Diminished
immunoreactive gastric inhibitory polypeptide response to a meal in newly diagnosed type I
(insulin-dependent) diabetics. J Clin Endocrinol Metab. 56, 1306-12.
12. Naitoh R, Miyawaki K, Harada N, Mizunoya W, Toyoda K, Fushiki T, Yamada Y, Seino Y, Inagaki N.(2008) Inhibition of GIP signaling modulates adiponectin levels under high-fat diet in mice. Biochem Biophys Res Commun. 376, 21-5.
13. Miyawaki K, Yamada Y, Yano H, Niwa H, Ban N, Ihara Y, Kubota A, Fujimoto S, Kajikawa M,
Kuroe A, Tsuda K, Hashimoto H, Yamashita T, Jomori T, Tashiro F, Miyazaki J, Seino Y.
(1999) Glucose intolerance caused by a defect in the entero-insular axis: a study in gastric
inhibitory polypeptide receptor knockout mice. Proc Natl Acad Sci U S A. 96, 14843-7.
14. Miyawaki K, Yamada Y, Ban N, Ihara Y, Tsukiyama K, Zhou H, Fujimoto S, Oku A, Tsuda K,
Toyokuni S, Hiai H, Mizunoya W, Fushiki T, Holst JJ, Makino M, Tashita A, Kobara Y, Tsubamoto
Y, Jinnouchi T, Jomori T, Seino Y.(2002) Inhibition of gastric inhibitory polypeptide signaling
prevents obesity. Nat Med. 8, 738-42.
15. Tsukiyama K, Yamada Y, Yamada C, Harada N, Kawasaki Y, Ogura M, Bessho K, Li M, Amizuka N, Sato M, Udagawa N, Takahashi N, Tanaka K, Oiso Y, Seino Y. (2006) Gastric inhibitory polypeptide as an endogenous factor promoting new bone formation after food ingestion. Mol Endocrinol. 20, 1644-51.
To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.