Human Aβ1-40(Amyloid Beta 1-40) ELISA Kit
人β淀粉样蛋白1-40(Aβ1-40)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-H0542
反应性: 人
检测范围: 15.63~1000 pg/mL
灵敏度: 9.38 pg/mL
用途
该试剂盒用于体外定量检测人serum, plasma and other biological fluids中Aβ1-40浓度
检测原理
本试剂盒采用双抗体夹心ELISA法。用抗人Aβ1-40抗体包被于酶标板上，实验时样品或标准品中的人Aβ1-40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Aβ1-40抗体和辣根过氧化物酶标记的亲和素。抗人Aβ1-40抗体与结合在包被抗体上的人Aβ1-40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，Aβ1-40浓度与OD450值之间呈正比，通过绘制标准曲线计算出样品中Aβ1-40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 | 15.63—1000 pg/mL |
| 灵敏度 | 9.38 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Aβ1-40，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 |
|---|---|---|---|
| 1000 | 2.395 2.429 |
2.412 | 2.33 |
| 500 | 1.589 1.623 |
1.606 | 1.524 |
| 250 | 0.966 0.934 |
0.95 | 0.868 |
| 125 | 0.464 0.502 |
0.483 | 0.401 |
| 62.5 | 0.261 0.249 |
0.255 | 0.173 |
| 31.25 | 0.183 0.181 |
0.182 | 0.1 |
| 15.63 | 0.132 0.136 |
0.134 | 0.052 |
| 0 | 0.075 0.089 |
0.082 | -- |
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) |
50.60 | 152.10 | 454.70 | 45.80 | 156.80 | 436.80 |
| Standard
deviation |
3.30 | 6.10 | 23.60 | 2.60 | 9.10 | 13.10 |
| C V (%) | 6.52 | 4.01 | 5.19 | 5.68 | 5.80 | 3.00 |
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) | 93-109 | 101 |
| EDTA plasma (n=5) | 85-96 | 91 |
| Cell culture media (n=5) | 92-108 | 100 |
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 (%) | 86-101 | 96-110 | 90-106 |
| Average (%) | 93 | 104 | 97 | |
| 1:4 | Range (%) | 99-116 | 80-92 | 95-111 |
| Average (%) | 106 | 87 | 102 | |
| 1:8 | Range (%) | 100-113 | 84-98 | 96-108 |
| Average (%) | 106 | 90 | 102 | |
| 1:16 | Range (%) | 97-110 | 81-92 | 97-109 |
| Average (%) | 105 | 87 | 103 |
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 |
客户发表文献
- 文献: Interneuron Accumulation of Phosphorylated tau Impairs Adult Hippocampal Neurogenesis by Suppressing GABAergic Transmission
样品类型: Cell culture supernatant - 文献: Modulating autophagic flux via ROS-responsive targeted micelles to restore neuronal proteostasis in Alzheimer's disease
样品类型: HT22 cell - 文献: Activation of PPARA-mediated autophagy reduces Alzheimer disease-like pathology and cognitive decline in a murine model
样品类型: Tissue homogenate,Cell culture supernatant,Cell lysate - 文献: Brain-neuron targeted nanoparticles for peptide synergy therapy at dual-target of Alzheimer's disease
样品类型: hippocampal tissues - 文献: Platelet biomarkers identifying mild cognitive impairment in type 2 diabetes patients
样品类型: plasma - 文献: CIP2A Causes Tau/APP Phosphorylation, Synaptopathy, and Memory Deficits in Alzheimer’s Disease
样品类型: Cell lysate - 文献: Far infrared light irradiation enhances Aβ clearance via increased exocytotic microglial ATP and ameliorates cognitive deficit in Alzheimer’s disease-like mice
样品类型: brain - 文献: AP2S1 regulates APP degradation through late endosome-lysosome fusion in cells and APP/PS1 mice
- 文献: Glycopeptide Nanofiber Platform for Aβ-Sialic Acid Interaction Analysis and Highly Sensitive Detection of Aβ
样品类型: serum - 文献: Periphery Biomarkers for Objective Diagnosis of Cognitive Decline in Type 2 Diabetes Patients
样品类型: plasma - 文献: A monomeric polysaccharide from Polygonatum sibiricum improves cognitive functions in a model of Alzheimer's disease by reshaping the gut microbiota
样品类型: hippocampus tissue - 文献: Periodontitis and systemic markers of neurodegeneration: A case–control study
样品类型: Serum - 文献: Alpha- and Gamma-Tocopherol Modulates the Amyloidogenic Pathway of Amyloid Precursor Protein in an in vitro Model of Alzheimer’s Disease: A Transcriptional Study
样品类型: SH-SY5Y cell - 文献: Purified oleocanthal and ligstroside protect against mitochondrial dysfunction in models of early Alzheimer's disease and brain ageing
样品类型: Cell lysate - 文献: Genistein Decreases APP/tau Phosphorylation and Ameliorates Aβ Overproduction Through Inhibiting CIP2A
样品类型: HEK-293T cells - 文献: The mitochondria-targeted small molecule SS31 delays progression of behavioral deficits by attenuating β-amyloid plaque formation and mitochondrial/synaptic deterioration in APP/PS1 mice
样品类型: hippocampus - 文献: Periodontitis is associated with systemic inflammation and vascular endothelial dysfunction in patients with lacunar infarct
样品类型: Serum - 文献: Influence of manganese exposure on cognitive function, plasma APP and Aβ levels in older men
样品类型: Plasma - 文献: Comparative Effects of Alpha- and Gamma-Tocopherol on Mitochondrial Functions in Alzheimer’s Disease In Vitro Model
样品类型: Cell culture medium - 文献: Resveratrol Abrogates Hypoxia-Induced Up-Regulation of Exosomal Amyloid-β Partially by Inhibiting CD147
样品类型: Tissue homogenate,Cell culture medium - 文献: Evaluation of the Expression of Amyloid Precursor Protein and the Ratio of Secreted Amyloid Beta 42 to Amyloid Beta 40 in SH-SY5Y Cells Stably Transfected with Wild-Type, Single-Mutant and Double-Mutant Forms of the APP Gene for the Study of Alzheimer’s D
样品类型: Cell culture supernatant
