1:预包被板: 96T/48T

2:酶标记抗体: (30倍浓缩)HRPIgG,亲合纯化 0.4mL x 1

3:标准品: -6 0.5mL x 2

4:EIA缓冲液: 含1% BSA, 0.05%吐温20 BPS 30mL x 1

5:标记抗体稀释液: 含1% BSA, 0.05%吐温20 BPS 12mL x 1

6:显色剂: TMB底物液 15mL x 1

7:终止液: 1N硫酸 12mL x 1

8:浓缩洗涤液: (40倍浓缩) 含1% BSA, 0.05%吐温20 BPS 50mL x 1

艾美捷血管紧张素ⅡELISA试剂盒操作流程：

(1)血管紧张素Ⅱ ELISA试剂盒待测样品孔中每孔加入待测样品100μl,每种样品设3个平行孔;设两个阴性对照孔,每孔加未处理组的细胞裂解液100μl;另设一个空白对照孔,加入纯细胞裂解液100μl。

(2)酶标板置4℃,包被过夜。

(3)洗板:吸干孔内反应液,仅用于科研用洗涤液过洗一遍(将洗涤液注满板孔后,即甩去),之后将洗涤液注满板孔,浸泡1-2分钟,间歇摇动。甩去孔内液体后在吸水纸上拍干。重复洗涤3-4次。

(4)阴性对照孔每孔加入PBS 50μl,样品孔及空白孔每孔加入1:500稀释的兔抗人AIF抗体工作液50μl。

(5)酶标板置37℃培养箱的湿盒内,孵育60min。

(6)洗板,同(4)。

(7)每孔加1:5000稀释的HRP-标记的山羊抗兔抗体工作液 100μl。

(8)酶标板置37℃培养箱的湿盒内,孵育60min。

(9)洗板,同(4)。

(10)每孔加TMB显色液 100μl,轻轻混匀10s,置37℃暗处反应15-20min。

(11)每孔加100μl 2mol/L H2SO4终止反应。

(12)分别测450nm 吸光值W1和630nm吸光值W2,终测得的OD值为两者之差(W1-W2),以减少由容器上的划痕或指印等造成的光干扰。

(13)数据处理:在得出标本(S)和阴性对照(N)的 OD值后,计算S/N值。S/N≥2.1为阳性判定标准。

血管紧张素ⅡELISA试剂盒文献参考：

Co-ingestion of whole eggs or egg whites with glucose protects against postprandial hyperglycaemia-induced oxidative stress ａnd dysregulated arginine metabolism in association with improved vascular endothelial function in prediabetic men: J.D. McDonald, et al.; Br. J. Nutr. 120, 901 (2018), Application(s): Human blood plasma；

Hydrogen sulfide attenuates renin angiotensin ａnd aldosterone pathological signaling to preserve kidney function ａnd improve exercise tolerance in heart: Z. Li, et al.; JACC Basic Transl. Sci. 3, 796 (2018), Application(s): ELISA using mouse kidney；

Hypertension enhances advanced atherosclerosis ａnd induces cardiac death in Watanabe heritable hyperlipidemic rabbits: B. Ning, et al.; Am. J. Pathol. 188, 2936 (2018)；

Post-translational modifications of eNOS augment nitric oxide availability ａnd facilitates hypoxia adaptation in Ladakhi women: Pooja, et al.; Nitric Oxide 78, 103 (2018), Application(s): ELISA using human plasma；

Prorenin independently causes hypertension ａnd renal ａnd cardiac fibrosis in cyp1a1-prorenin transgenic rats: G. Zhou, et al.; Clin. Sci. (Lond.) 132, 1345 (2018)；

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