1:預(yù)包被板: 96T/48T

2:酶標(biāo)記抗體: (30倍濃縮)HRPIgG,親合純化 0.4mL x 1

3:標(biāo)準(zhǔn)品: -6 0.5mL x 2

4:EIA緩沖液: 含1% BSA, 0.05%吐溫20 BPS 30mL x 1

5:標(biāo)記抗體稀釋液: 含1% BSA, 0.05%吐溫20 BPS 12mL x 1

6:顯色劑: TMB底物液 15mL x 1

7:終止液: 1N硫酸 12mL x 1

8:濃縮洗滌液: (40倍濃縮) 含1% BSA, 0.05%吐溫20 BPS 50mL x 1

艾美捷血管緊張素ⅡELISA試劑盒操作流程：

(1)血管緊張素Ⅱ ELISA試劑盒待測(cè)樣品孔中每孔加入待測(cè)樣品100μl,每種樣品設(shè)3個(gè)平行孔;設(shè)兩個(gè)陰性對(duì)照孔,每孔加未處理組的細(xì)胞裂解液100μl;另設(shè)一個(gè)空白對(duì)照孔,加入純細(xì)胞裂解液100μl。

(2)酶標(biāo)板置4℃,包被過(guò)夜。

(3)洗板:吸干孔內(nèi)反應(yīng)液,僅用于科研用洗滌液過(guò)洗一遍(將洗滌液注滿板孔后,即甩去),之后將洗滌液注滿板孔,浸泡1-2分鐘,間歇搖動(dòng)。甩去孔內(nèi)液體后在吸水紙上拍干。重復(fù)洗滌3-4次。

(4)陰性對(duì)照孔每孔加入PBS 50μl,樣品孔及空白孔每孔加入1:500稀釋的兔抗人AIF抗體工作液50μl。

(5)酶標(biāo)板置37℃培養(yǎng)箱的濕盒內(nèi),孵育60min。

(6)洗板,同(4)。

(7)每孔加1:5000稀釋的HRP-標(biāo)記的山羊抗兔抗體工作液 100μl。

(8)酶標(biāo)板置37℃培養(yǎng)箱的濕盒內(nèi),孵育60min。

(9)洗板,同(4)。

(10)每孔加TMB顯色液 100μl,輕輕混勻10s,置37℃暗處反應(yīng)15-20min。

(11)每孔加100μl 2mol/L H2SO4終止反應(yīng)。

(12)分別測(cè)450nm 吸光值W1和630nm吸光值W2,終測(cè)得的OD值為兩者之差(W1-W2),以減少由容器上的劃痕或指印等造成的光干擾。

(13)數(shù)據(jù)處理:在得出標(biāo)本(S)和陰性對(duì)照(N)的 OD值后,計(jì)算S/N值。S/N≥2.1為陽(yáng)性判定標(biāo)準(zhǔn)。

血管緊張素ⅡELISA試劑盒文獻(xiàn)參考：

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)；

來(lái)源：https://www.amyjet.com/products/ADI-900-204.shtml