小鼠卵清蛋白特异性免疫球蛋白E（OVA-IgE） ELISA试剂盒
IgE（Immunoglobulin E，免疫球蛋白E）是第五个被发现的免疫球蛋白，由5个结构域（VH、CHε 1～4）构成的两个Hε链和两个L链中组成的IgE，其分子量约190000，电泳实验中向γ1域移动。IgE的代谢半衰期约3天，正常人血清中的IgE浓度非常低，约300 ng/mL。但在寄生虫感染和枯草热时浓度会升高。与过敏性相关的IgE被称作反应素。因接触过敏原导致反应素含量升高，反应素会在Fc域与存在于皮肤、呼吸道、消化脏器中嗜碱性粒细胞和肥大细胞的FcεR1受体结合，引起细胞过敏。在结合过敏原后细胞发生脱粒现象，组胺，五羟色胺，蛋白酶，肝素，趋化因子，前列腺素，白三烯等被投放，经过支气管收缩和黏膜水肿，分泌亢进，从而诱发支气管哮喘，部分荨麻疹，过敏性鼻炎，过敏性反应等I型过敏性反应。
本试剂盒是以OVA（卵清蛋白）作为免疫的抗原，在一个简化的反应系统中通过测定鼠抗OVA- IgE抗体值来进行小鼠免疫系统检测的试剂盒。
◆特点
● 测定时间短（总反应时间：1小时50分钟）。
● 微量样本即可测定。
● 使用无害的防腐剂。
● 全部试剂为溶液即用类型。
● 高测定精度和高重复性。
● 操作简便，无需进行特殊前处理。
◆试剂盒组成
组成品 | 状态 | 包装 |
OVA包被96孔板（干燥板） | 清洗后使用 | 96 wells（8×12）／1个 |
标准溶液（Anti OVA-IgE:1,200 U/mL）（单抗） | 稀释后使用 | 100 μL/1瓶 |
缓冲液 | 直接使用 | 60 mL/1瓶 |
生物素结合抗小鼠IgE抗体（单抗） | 稀释后使用 | 200 μL/1瓶 |
过氧化物酶·抗生素结合物 | 稀释后使用 | 200 μL/1瓶 |
显色液（TMB） | 直接使用 | 12 mL/1瓶 |
终止液（1M H2SO4）※小心轻放 | 直接使用 | 12 mL/1瓶 |
浓缩清洗液（10×） | 稀释后使用 | 100 mL/1瓶 |
孔板密封膜 | 3个 | |
产品说明书 | 1本 |
◆样本信息
● 小鼠血清·血浆
● 10 μL/well（稀释样本）
※ 样本需要用试剂盒附带的缓冲液调至标准曲线范围内。
※ 样本必须稀释10倍以上。
◆测定范围
1.88～120 U/mL（标准曲线范围）
（本试剂盒中1 U/mL定义为抗原结合常数（Ka）为2.0×108 M-1的抗体1.3 ng/mL）
◆实验数据
精度测试（组内变异）
样本 | A | B |
1 | 70.7 | 19.1 |
2 | 71.0 | 18.7 |
3 | 77.1 | 19.6 |
4 | 74.3 | 19.7 |
5 | 72.9 | 18.9 |
Mean | 73.2 | 19.2 |
SD | 2.6 | 0.42 |
CV（%） | 3.6 | 2.2 |
单位：U/mL
重复性测试（组间变异）
测定日/样本 | C | D | E |
0天 | 60.0 | 15.0 | 3.75 |
1天 | 59.1 | 15.0 | 3.75 |
2天 | 58.1 | 14.7 | 3.65 |
3天 | 63.6 | 16.0 | 3.48 |
Mean | 60.2 | 15.2 | 3.66 |
SD | 2.4 | 0.57 | 0.13 |
CV（%） | 4.0 | 3.7 | 3.4 |
单位：U/mL
加标回收测试
样本F
添加量 | 实测值 | 回收量 | 回收率（%） |
0.00 | 6.93 | — | — |
5.35 | 12.2 | 5.27 | 98.5 |
10.7 | 17.8 | 10.9 | 102 |
17.1 | 25.1 | 18.2 | 106 |
单位：U/mL，n=3
样本G
添加量 | 实测值 | 回收量 | 回收率（%） |
0.00 | 40.5 | — | — |
30.8 | 70.4 | 29.5 | 95.8 |
35.9 | 77.1 | 36.6 | 102 |
53.9 | 94.9 | 54.4 | 101 |
单位：U/mL，n=3
稀释直线性测试
2个血清样本连续用稀释缓冲液稀释3个梯度测定结果，直线回归值R2=0.9987~0.9999
参考文献
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| 产品编号 | 产品名称 | 产品规格 | 产品等级 | 备注 |
| 639-07651 | （AKRIE-030）小鼠卵清蛋白特异性免疫球蛋白E(OVA-IgE) ELISA试剂盒 LBIS® OVA-IgE Mouse |
96 tests | - | - |
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