PGE2(Prostaglandin E2) ELISA Kit

前列腺素E2(PGE2)酶联免疫吸附测定试剂盒

规格: / 96T / 48T / 96T*5
价格: / ¥2400 / ¥1650 / ¥10200

货号:E-EL-0034c

货期:3个工作日+顺丰周期

反应性: 通用

检测范围: 31.25~2000 pg/mL

灵敏度: 18.75 pg/mL

  • 详情
  • 文献(33)
  • 用途

    该试剂盒用于体外定量检测血清、血浆或其他相关生物液体中天然PGE2浓度。

    检测原理

    本试剂盒采用竞争ELISA法。用PGE2抗原包被于酶标板上,实验时样品或标准品中的PGE2与包被的PGE2竞争生物素标记的抗PGE2单抗上的结合位点,游离的成分被洗去。加入辣根过氧化物酶标记的亲和素,生物素与亲和素特异性结合而形成免疫复合物,游离的成分被洗去。加入显色底物(TMB),TMB在辣根过氧化物酶的催化下呈现蓝色,加终止液后变成黄色。用酶标仪在450nm波长处测OD值,PGE2浓度与OD450值之间呈反比,通过绘制标准曲线计算出样品中PGE2的浓度。

    反应类型 竞争法
    规格 96T
    反应时间 2.0h
    反应性 通用
    检测方法 Colormetric
    检测范围 31.25-2000 pg/mL
    灵敏度 18.75 pg/mL
    样本体积 50μL
    样本类型 血清、血浆或其他相关生物液体

    特异性

    可检测样本中的PGE2,且与其它相关蛋白无明显交叉反应。

    重复性

    板内,板间变异系数均<10%。

    典型数据

    由于实验操作条件的不同(如操作者、移液技术、洗板技术和稳定条件等),标准曲线的OD值会有所差异。以下数据和曲线仅供参考,实验者需根据自己的实验建立标准曲线。

    (pg/mL) O.D Average
    2000 0.381
    0.393
    0.387
    1000 0.489
    0.517
    0.503
    500 0.702
    0.696
    0.699
    250 0.974
    1.002
    0.988
    125 1.341
    1.331
    1.336
    62.5 1.674
    1.662
    1.668
    31.25 1.913
    1.925
    1.919
    0 2.261
    2.269
    2.265

    精密度

    板内精密度:低浓度样本,中浓度样本和高浓度样本分别在1块板子上检测20次。
    板间精密度:低浓度样本,中浓度样本和高浓度样本分别在3块板子上检测20次。

    Intra-assay Precision Inter-assay Precision
    Sample 1 2 3 1 2 3
    n 20 20 20 20 20 20
    Mean
    (pg/mL)
    101.30 236.80 779.30 96.00 259.90 728.60
    Standard
    deviation
    6.60 11.60 42.10 4.90 13.80 31.30
    C V (%) 6.52 4.90 5.40 5.10 5.31 4.30

    回收率

    分别往5个不同样本中添加已知浓度的目标蛋白,做回收实验,得出回收率范围和平均回收率。

    Sample Type Range (%) Average Recovery (%)
    Serum (n=5) 92-109 99
    EDTA plasma (n=5) 87-99 94
    Cell culture media (n=5) 86-96 91

    线性

    分别往5个样本中添加已知浓度的目标蛋白,做回收实验,得出回收率范围及平均回收率。将5个样本分别稀释2倍,4倍,8倍,16倍做回收实验,得出回收率范围及平均回收率。

    Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
    1:2 Range (%) 91-104 89-101
    Average (%) 103 98 96
    1:4 Range (%) 85-95 89-101 101-117
    Average (%) 90 95 107
    1:8 Range (%) 87-100 92-104 96-111
    Average (%) 92 97 102
    1:16 Range (%) 88-104 93-105 99-114
    Average (%) 95 98 106

    试剂盒组成及保存

    未拆封的试剂盒可在4℃保存一周;如果一周以后才使用试剂盒,请拆开试剂盒按照下表中的条件分别保存各组分。

    中文名称 英文名称 规格 保存条件
    ELISA酶标板(可拆卸) Micro ELISA Plate(Dismountable) 8 孔×12 条 -20℃,可存放6个月
    冻干标准品 Reference Standard 2支
    浓缩生物素化抗体(100×) Concentrated Biotinylated Detection Ab 1支 120 μL
    浓缩HRP酶结合物(100×) Concentrated HRP Conjugate 1支 120 μL -20℃(避光),可存放6个月
    标准品&样品稀释液 Reference Standard & Sample Diluent 1瓶 20 mL 4℃.可存放6个月
    生物素化抗体稀释液 Biotinylated Detection Ab Diluent 1瓶 14 mL
    酶结合物稀释液 HRP Conjugate Diluent 1瓶 14 mL
    浓缩洗涤液(25×) Concentrated Wash Buffer (25×) 1瓶 30 mL
    底物溶液(TMB) Substrate Reagent 1瓶 10 mL 4℃(避光)
    反应终止液 Stop Solution 1瓶 10 mL 4℃
    封板覆膜 Plate Sealer 5 张
    产品说明书 Product Description 1 份
    质检报告 Certificate of Analysis 1 份

    说明:所有试剂瓶盖须旋紧以防止蒸发和微生物的污染。
    试剂体积以实际发货版说明书为准。相关试剂在分装时会比标签上标明的体积稍多一些,请在使用时量取而非直接倒出。

    试验所需自备物品

    • 1.酶标仪(450nm波长滤光片)
    • 2.高精度移液器,EP管及一次性吸头:0.5-10 μL, 2-20 μL, 20-200 μL, 200-1000 μL
    • 3.37℃恒温箱,双蒸水或去离子水
    • 4.吸水纸

    操作步骤

    1. 对应板孔中加入50μL标准品工作液或样本后,立即每孔加入50μL生物素化抗体工作液,37℃孵育90分钟
    2. 弃掉板内液体,洗板3次
    3. 每孔加入100μL HRP酶结合物工作液,37℃孵育30分钟,弃掉板内液体,洗板5次
    4. 每孔加入90μL底物溶液,37℃孵育15分钟
    5. 每孔加入50μL终止液
    6. 立即在450nm波长下读数,处理数据

    客户发表文献

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