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Amplite™ Fluorimetric Myeloperoxidase Assay Kit
/
详见说明
齐源生物
详见说明
200次
样品分析
表1. 96孔固体黑色微孔板中MPO标准品和测试样品的布局。 MPO =髓过氧化物酶标准品(MPO1-MPO7,0.01至10mU / mL); BL =空白对照; TS =测试样品。
BL | BL | TS | TS |
MPO1 | MPO1 | ... | ... |
MPO2 | MPO2 | ... | ... |
MPO3 | MPO3 | ||
MPO4 | MPO4 | ||
MPO5 | MPO5 | ||
MPO6 | MPO6 | ||
MPO7 | MPO7 |
表2.每个孔的试剂组成。 注意,由于Amplite Red底物(非荧光产物)的过度氧化,高浓度的MPO可能导致荧光信号降低。
孔 | 容积 | 试剂 |
MPO1-MPO7 | 50ul | 连续稀释(0.01至10 mU / mL) |
BL | 50ul | 分析缓冲液(组分B) |
TS | 50ul | 测试样本 |
1.根据表1和2中提供的布局制备髓过氧化物酶标准品(MPO),空白对照(BL)和测试样品(TS)。对于384孔板,每孔使用25μL试剂代替50μL。
2.向髓过氧化物酶标准品,空白对照和测试样品的每个孔中加入50μLMPO工作溶液,使总MPO测定体积为100μL/孔。 对于384孔板,在每个孔中加入25μLMPO工作溶液,总体积为50μL/孔。
3.在室温下孵育反应30至60分钟,避光。
4.用荧光板读数器在激发= 530-570nm,发射= 590-600nm(最佳Ex / Em = 540 / 590nm,截止= 570nm)监测荧光强度。 注意:也可以将板的内容物转移到白色透明底板上,并用吸光度酶标仪在576±5nm的波长下读数。 与荧光读数相比,吸收检测具有更低的灵敏度。
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