Human MCP-1(Monocyte Chemotactic Protein 1) ELISA Kit
人单核细胞趋化蛋白1(MCP-1)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-H6005
反应性: 人
检测范围: 62.50~4000 pg/mL
灵敏度: 37.50 pg/mL
用途
该试剂盒用于体外定量检测人serum, plasma and other biological fluids中MCP-1浓度
检测原理
本试剂盒采用双抗体夹心ELISA法。用抗人MCP-1抗体包被于酶标板上,实验时样品或标准品中的人MCP-1会与包被抗体结合,游离的成分被洗去。依次加入生物素化的抗人MCP-1抗体和辣根过氧化物酶标记的亲和素。抗人MCP-1抗体与结合在包被抗体上的人MCP-1结合、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,MCP-1浓度与OD450值之间呈正比,通过绘制标准曲线计算出样品中MCP-1的浓度。
反应类型 | 夹心法 |
规格 | 96T |
反应时间 | 3.5h |
反应性 | 人 |
检测方法 | Colormetric |
检测范围 | 62.50—4000 pg/mL |
灵敏度 | 37.50 pg/mL |
样本体积 | 100μL |
样本类型 | serum, plasma and other biological fluids |
特异性
可检测样本中的人MCP-1,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 | Corrected |
---|---|---|---|
4000 | 2.552 2.507 |
2.53 | 2.469 |
2000 | 1.634 1.645 |
1.64 | 1.579 |
1000 | 0.957 0.946 |
0.952 | 0.891 |
500 | 0.487 0.506 |
0.497 | 0.436 |
250 | 0.275 0.268 |
0.272 | 0.211 |
125 | 0.19 0.19 |
0.19 | 0.129 |
62.50 | 0.129 0.113 |
0.121 | 0.06 |
0 | 0.057 0.064 |
0.061 | -- |
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) |
198.91 | 488.33 | 1680.55 | 164.36 | 433.06 | 1730.78 |
Standard
deviation |
10.01 | 24.12 | 90.58 | 9.45 | 20.74 | 76.85 |
C V (%) | 5.03 | 4.94 | 5.39 | 5.75 | 4.79 | 4.44 |
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) | 97-108 | 101 |
EDTA plasma (n=5) | 89-99 | 94 |
Cell culture media (n=5) | 87-95 | 92 |
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 (%) | 98-110 | 89-102 | 91-104 |
Average (%) | 103 | 96 | 98 | |
1:4 | Range (%) | 87-98 | 90-101 | 94-100 |
Average (%) | 92 | 94 | 97 | |
1:8 | Range (%) | 90-100 | 94-103 | 101-111 |
Average (%) | 94 | 99 | 105 | |
1:16 | Range (%) | 89-99 | 84-96 | 89-101 |
Average (%) | 96 | 92 | 97 |
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100μL标准品工作液或样本,37℃孵育90分钟 |
2.弃掉板内液体后,立即加入100μL生物素化抗体工作液,37℃孵育60分钟 | |
3.弃掉板内液体,洗板3次 | |
4.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 | |
5.每孔加入90μL底物溶液,37℃孵育15分钟 | |
6.每孔加入50μL终止液 | |
7.立即在450nm波长下读数,处理数据 |
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