MS Mouse VEGF-A(Vascular Endothelial Cell Growth Factor A) ELISA Kit
MS-小鼠血管内皮细胞生长因子(VEGF-A)酶联免疫吸附测定试剂盒
规格: | / 96T / 48T / 96T*5 |
价格: | / ¥3080 / ¥2180 / ¥13860 |
货号:E-MSEL-M0005
反应性: 小鼠
检测范围: 7.81~500 pg/mL
灵敏度: 3.21 pg/mL
MS系列
MS为Mini Sample的缩写。MS系列试剂盒的上样量仅为传统夹心法ELISA试剂盒的1/4,能有效解决实验样本体积不够的问题。Elabscience®自主开发的新技术,旨在帮助您以更高效的方式进行科学研究。
用途
该试剂盒用于体外定量检测小鼠serum, plasma and other biological fluids中VEGF-A浓度
检测原理
本试剂盒采用双抗体夹心ELISA法。用抗小鼠VEGF-A抗体包被于酶标板上,实验时样品或标准品中的小鼠VEGF-A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小鼠VEGF-A抗体和辣根过氧化物酶标记的亲和素。抗小鼠VEGF-A抗体与结合在包被抗体上的小鼠VEGF-A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,VEGF-A浓度与OD450值之间呈正比,通过绘制标准曲线计算出样品中VEGF-A的浓度。
反应类型 | 夹心法 |
规格 | 96T |
反应时间 | 3.5h |
反应性 | 小鼠 |
检测方法 | Colormetric |
检测范围 | 7.81—500 pg/mL |
灵敏度 | 3.21 pg/mL |
样本体积 | 25μL |
样本类型 | serum, plasma and other biological fluids |
特异性
可检测样本中的小鼠VEGF-A,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 |
---|---|---|---|
500 | 1.962 1.963 |
1.962 | 1.875 |
250 | 1.237 1.226 |
1.231 | 1.144 |
125 | 0.701 0.697 |
0.699 | 0.611 |
62.5 | 0.402 0.393 |
0.397 | 0.310 |
31.25 | 0.255 0.243 |
0.249 | 0.161 |
15.63 | 0.166 0.161 |
0.163 | 0.076 |
7.81 | 0.13 0.127 |
0.128 | 0.041 |
0 | 0.085 0.089 |
0.086 | -- |
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) |
22.41 | 58.49 | 215.21 | 20.44 | 50.95 | 220.39 |
Standard
deviation |
1.32 | 2.81 | 10.16 | 1.03 | 2.39 | 10.01 |
C V (%) | 5.91 | 4.81 | 4.72 | 5.06 | 4.69 | 4.54 |
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) | 99-105 | 102 |
EDTA plasma (n=5) | 93-100 | 96 |
Cell culture media (n=5) | 87-98 | 90 |
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 (%) | 93-103 | 85-98 | 91-103 |
Average (%) | 99 | 91 | 99 | |
1:4 | Range (%) | 94-102 | 98-107 | 95-108 |
Average (%) | 97 | 101 | 101 | |
1:8 | Range (%) | 88-95 | 85-95 | 95-108 |
Average (%) | 91 | 90 | 103 | |
1:16 | Range (%) | 87-97 | 93-104 | 87-94 |
Average (%) | 92 | 99 | 90 |
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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