LBIS® GLP-1(active) ELISA KIT
LBIS® 胰高血糖素样肽-1(GLP-1)(活性) ELISA 试剂盒
胰高血糖素样肽-1(Glucagon-like peptide-1,GLP-1)是胰高血糖素前驱体的一部分。胰高血糖素前驱体于胰脏、小肠下部以及下丘脑中表达。该前驱体的构造中含有与糖代谢有关的各种各样的生理活性物质(胰高血糖素,肠高血糖素,胃泌酸调节素,GLP-1,GLP-2)的氨基酸序列。根据表达部位加工酶的特异性,胰脏主要分泌胰高血糖素,而小肠下部主要分泌肠高血糖素和胃泌酸调节素。GLP-1 和 GLP-2 则存在于胰高血糖素前驱体后半的结构中。GLP-1由37个氨基酸组成,有2种生物活性形式,分别为 GLP-1(7-37)和GLP-1(7-36)酰胺。两者都存在于小肠下部、胰脏和下丘脑中,GLP-1(7-36)酰胺在下丘脑中占免疫反应 GLP-1(IR-GLP-1)总量的 55-94%,在小肠中占 27-73%。但在胰脏中只有极少量存在。大部分哺乳类(如人类、大鼠、小鼠、牛、猪、狗等)的 GLP-1 结构相似。
GLP-1: hdeferhaegtftsdvssylegqaakefiawlvkgrg
GLP 1(7-37): haegtftsdvssylegqaakefiawlvkgrg
GLP 1(7-36) amide: haegtftsdvssylegqaakefiawlvkgr-NH2
GLP-1 与小肠上部分泌的 GIP 统称为肠促胰素。该类激素是葡萄糖浓度依赖性方式促进胰岛素分泌。同时具有抑制胃肠道蠕动和胃液分泌、抑制胰高血糖素的释放、促进生长抑素的分泌、使食欲减退,促进肠上皮细胞生长、以及外周组织促进非胰岛素依赖性的葡萄糖的消耗,并促进细胞的生长的作用。有报告指出该类激素与垂体激素的分泌也有关系。
GLP-1(7-36)酰胺在生物体内代谢迅速,DPP-IV(dipeptidyl peptidase IV)会使其失去N-末端的两个氨基酸变为 GLP-1(9-36)酰胺,GLP-1(7-37)变为 GLP-1(9-37)后会失去活性。有报告指出,体外实验中,在犬的血浆中 GLP-1(7-36)酰胺的半衰期是为 61±9 分,GLP-1(7-37)为 132±16 分。因此 GLP-1 的测定,取样的时候有必要使用 DPP-IV 抑制剂。
此外,肠促胰素中的 GIP 则是有力促进 GLP-1 分泌的激素。回肠中 GLP-1 的分泌不是食物直接刺激肠道而产生的,而是由于胆碱能和肽类的刺激所产生的。
◆特点
● 短时间测定(完全反应时间:5小时)
● 微量样品(标准操作用量:10 μL)可测
● 使用对环境无害的防腐剂
● 全部试剂均为液体,可直接使用
● 精密的测定精度和高再现性
◆构成
组成 | 状态 | 容量 |
(A) 抗体固相化 96 孔板 | 洗净后使用 | 96 wells(8×12)/1 块 |
(B) GLP-1标准溶液(500 pg/mL) | 稀释后使用 | 200 μL/1 瓶 |
(C) 缓冲液 | 即用 | 60 mL/1 瓶 |
(D)生物素结合抗GLP-1抗体 | 稀释后使用 | 100 μL/1 瓶 |
(E) 过氧化物・抗生物素蛋白结合物 | 稀释后使用 | 100 μL/1 瓶 |
(F) 显色液(TMB) | 即用 | 12 mL/1 瓶 |
(H) 反应终止液(1M H2SO4)※小心轻放 | 即用 | 12 mL/1 瓶 |
( I ) 浓缩洗净液(10×) | 稀释后使用 | 100 mL/1 瓶 |
封板膜 | 4 张 | |
使用说明书 | 1 份 |
◆交叉反应
※交差率是 1,000 pg/mL 浓度时的数据
动物类型 | 对象物质 | 反应性和反应率(%) |
Mouse/Rat | GLP-1(7-36)amide | 100 |
GLP-1(7-37) | < 0.1 | |
GLP-1(1-37) | ― | |
GLP-1(9-36)amide | ― | |
GLP-2 | ― | |
Glucagon(1-29) | ― | |
Insulin | ― | |
Secretin | ― | |
GIP | ― | |
VIP | ― | |
GRF | ― | |
Bovine | Glucagon(1-29) | ― |
VIP | ― | |
Porcine | Glucagon(1-29) | ― |
VIP | ― |
―:不存在交叉反应
◆样品信息
小鼠和大鼠的血清及血浆
10 μL/well(标准操作方法)
※测量中由于酶(DPP-IV 等)的影响,采血时请注意防止 GLP-1(7-36)酰胺的分解,再使用。
◆测定范围
1.56~50.0 pg/mL 【0.47~15.16 pmol/L(分子量3298)】(标准曲线范围)
7.8~250 pg/mL(样品量 10 μL)
3.9~125 pg/mL(样品量 20 μL)
◆Validation data
精度测试(组内变异)
样品 | A | B |
1 | 23.7 | 6.44 |
2 | 23.2 | 5.97 |
3 | 23.4 | 6.39 |
4 | 24.0 | 5.87 |
5 | 24.1 | 6.44 |
mean | 23.7 | 6.22 |
SD | 0.35 | 0.28 |
CV(%) | 1.5 | 4.5 |
单位:pg/mL
重复性测试(组间变异)
测量日/样品 | E | F |
第0天 | 25.1 | 6.31 |
第1天 | 25.1 | 6.16 |
第2天 | 25.0 | 6.24 |
第3天 | 25.0 | 6.37 |
mean | 25.0 | 6.27 |
SD | 0.03 | 0.09 |
CV(%) | 0.13 | 1.4 |
单位:pg/mL n=4
加标回收测试
样品C
添加量 | 实测值 | 回收量 | 回收率(%) |
0.00 | 3.93 | – | – |
3.26 | 7.28 | 3.35 | 103 |
6.51 | 10.3 | 6.37 | 97.8 |
8.14 | 12.1 | 8.17 | 100 |
样品D
添加量 | 实测值 | 回收量 | 回收率(%) |
0.00 | 11.8 | – | – |
7.16 | 19.1 | 7.30 | 102 |
14.3 | 25.5 | 13.7 | 95.8 |
21.5 | 32.4 | 20.6 | 95.8 |
稀释直线性测试
用稀释缓冲液分三次连续稀释2个血清样品的结果,直线回归方程的R2在0.997~0.9999之间。
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产品编号 | 产品名称 | 产品规格 | 产品等级 | 备注 |
637-15129 | (AKMGP-011) LBIS® GLP-1(Active) ELISA Kit LBIS® 胰高血糖素样肽-1(GLP-1)(活性) ELISA试剂盒 |
96 tests | - | - |
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