YK250 Human GIP (Active) ELISA FOR RESEARCH LABORATORY USE ONLY 418-0011 2480-1 FAX: 0544-22-2770 TEL: 0544-22-2771 Website: www.yanaihara.co.jp E-mail: ask@yanaihara.co.jp
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YK250 Human GIP (Active) ELISA. GIP (glucose-dependent insulinotropic polypeptide)glp-1 (glucagon-like peptide-1)2 GIP 1970 Brown 42 GIP 2) GIP 3,4) 18 34 2 Takeda GIP GIP VIP GIP K GIP GIP GIP GIP GIP (1-42) DPP-4 GIP (3-42) DPP GIP GIP GIP (3-42) GIP (1-42) YK250 Human GIP (Active) ELISA GIP (1-42) 3.9250 pg/ml (0.7850.2 pm) 1) 40 duplicate 2) 3.5 3) 4) 50 µl 5) 8 6) 7) 8) 28 24 2
. GIP (1-42) GIP (1-42) GIP (1-42) 98% human GIP (1-42) human GIP (3-42) glucagon human GLP-2 GLP-1 (7-36)NH 2 GLP-1 (9-36)NH 2 96 GIP (1-42) HRP GIP (1-42) HRP GIP (1-42) 3
. 1. 2. 3. 4. 5. 6. 7. 8. 4
. 1. 25 µl1 ml8 12 2. 450 nm 3.0 3. 4. 5. 6. 1000 ml 7. 1. 1 ml 500 pg/ml 0.2 ml 0.2 ml 250 pg/ml 125, 62.5, 31.3, 15.6, 7.8, 3.9 pg/ml 0 pg/ml 3.9 pg/ml250 pg/ml (0.78 pm50.2 pm) 3.9 pg/ml 3.9 pg/ml 2 2 pg/ml 2. 50 ml 950 3. 1. 2030 2. 350 µl 2 3 3. 50 µl (0, 3.9, 7.8, 15.6, 31.3, 62.5, 125, 250 5
pg/ml) 50 µl 4. 2 100 rpm 5. 2. 4 6. 100 µl 7. 1 100 rpm 8. 1 9. 2. 4 10. 100 µl 30 11. 100 µl 12. 450 nm/620 nm 13. 5or 4 Parameter GIP GIP GIP 6
. 1. EDTA-2Na DPP- inhibitor0.01ml/ml (Catalog No. DPP4 MILLIPORE) BD TM P800 GLP-1, GIP, Glucagon, Ghrelin 30 2. 30 2 3. 4. 5. 250 pg/ml 6. 100 rpm 7. 8. - 9. 10. 11. 12. 7
VI. 10.00 Typical standard curve O.D. (450 / 620 nm) 1.00 0.10 0.01 1 10 100 1000 Human GIP (1-42) (pg/ml) A Added GIP (1-42) 0 14.3 5 18.1 19.3 93.8 25 37.6 39.3 95.7 100 111.8 114.3 97.8 Recovery B Added GIP (1-42) Recovery 0 26.7 5 32.7 31.7 103.2 25 46.5 51.7 89.9 100 110.5 126.7 87.2 8
C Added GIP (1-42) 0 9.4 5 12.9 14.4 89.6 25 30.2 34.4 87.8 100 108.1 109.4 98.8 Recovery D Added GIP (1-42) Recovery 0 22.1 5 27.2 27.1 100.4 25 47.1 47.1 100.0 100 113.8 122.1 93.2 E Added GIP (1-42) Recovery 0 9.3 5 13.4 14.3 93.7 25 32.8 34.3 95.6 100 109.9 109.3 100.5 A B C X1 72.1 72.1 X2 36.4 36.1 100.8 X4 19.4 18.0 107.8 X8 10.0 9.0 111.1 % of X1 46.3 46.3 X2 24.8 23.1 107.3 X4 12.8 11.6 110.6 X8 7.6 5.8 131.0 % of X1 30.1 30.1 X2 14.7 15.0 98.0 X4 8.0 7.5 106.7 % of 9
D X1 20.9 20.9 X2 11.1 10.5 105.7 X4 6.2 5.2 119.2 % of E X1 30.2 30.2 X2 15.9 15.1 105.3 X4 8.9 7.6 117.1 % of GIP (1-42) A Concentration Mean X1 Over X2 Over X4 171.7 686.8 X8 82.5 660.2 673.5 B Concentration Mean X1 Over X2 Over X4 234.5 938.0 X8 123.6 989.0 963.5 C Concentration Mean X1 Over X2 Over X4 Over X8 206.3 1650.2 1650.2 10
D Concentration Mean X1 Over X2 Over X4 Over X8 128.2 1025.5 1025.5 E Concentration Mean X1 Over X2 Over X4 165.8 663.1 X8 88.8 710.4 686.8 F Concentration Mean X1 Over X2 166.5 332.9 X4 82.3 329.1 X8 40.4 322.9 328.3 G Concentration Mean X1 Over X2 240.8 481.6 X4 123.6 494.3 X8 64.5 515.7 497.2 % GIP (1-42) (Human) 100 GIP (3-42) (Human) <0.1 Glucagon <0.1 Human GLP-2 <0.1 GLP-1 (7-36) NH 2 <0.1 GLP-1 (9-36) NH 2 <0.1 11
CV 5.26.5 CV 2.65.6 3.9 ~ 250 pg/ml(0.7850.2 pm) 28 24 96 1. Brown,J.C., Mutt, V. and Pedersen,R.A. (1970) Further purification of a polypeptide demonstrating enterogastrone activity. J.Physiol. 209, 57-64 2. Moody, A. J., Thim, L. & Valverde, I. (1984) The isolation and sequencing of human gastric inhibitory peptide(gip). FEBS Lett. 172,142-148. 3. Jörnvall 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. 4. Moody,A.J., Damm Jorgensen, K.and Thim, L.(1981)Diabetologia 21, 306, abstr. 5. 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. 6. Pederson, R.A. (1994) in Gut Peptides: Biochemistry and Physiology, eds, Walsh, J.H.& Dockray, G.J. (Raven, New York), pp,217-260 7. 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
8. 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. 9. 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. 10. 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. 11. 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. 418-0011 2480-1 FAX:0544-22-2770TEL:0544-22-2771 www.yanaihara.co.jpask@yanaihara.co.jp 2014 12 25 13