MS Mouse LEP(Leptin) ELISA Kit
MS-小鼠瘦素(LEP)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-M0033
反应性: 小鼠
检测范围: 0.63~40 ng/mL
灵敏度: 0.38 ng/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LEP浓度
检测原理
本试剂盒采用双抗体夹心ELISA法。用抗小鼠LEP抗体包被于酶标板上，实验时样品或标准品中的小鼠LEP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LEP抗体和辣根过氧化物酶标记的亲和素。抗小鼠LEP抗体与结合在包被抗体上的小鼠LEP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，LEP浓度与OD450值之间呈正比，通过绘制标准曲线计算出样品中LEP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 | 0.63—40 ng/mL |
| 灵敏度 | 0.38 ng/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LEP，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。
| (ng/mL) | O.D | Average | Corrected |
|---|---|---|---|
| 40 | 2.491 2.549 |
2.52 | 2.442 |
| 20 | 1.773 1.817 |
1.795 | 1.717 |
| 10 | 1.049 1.013 |
1.031 | 0.953 |
| 5 | 0.505 0.541 |
0.523 | 0.445 |
| 2.5 | 0.322 0.3 |
0.311 | 0.233 |
| 1.25 | 0.196 0.188 |
0.192 | 0.114 |
| 0.63 | 0.129 0.135 |
0.132 | 0.054 |
| 0 | 0.069 0.086 |
0.078 | -- |
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 (ng/mL) |
2.13 | 5.51 | 15.51 | 2.27 | 5.09 | 16.44 |
| Standard
deviation |
0.11 | 0.24 | 0.79 | 0.13 | 0.28 | 0.61 |
| C V (%) | 5.15 | 4.38 | 5.12 | 5.93 | 5.59 | 3.73 |
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) | 93-108 | 100 |
| EDTA plasma (n=5) | 93-108 | 101 |
| Cell culture media (n=5) | 90-105 | 96 |
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 (%) | 94-109 | 88-100 | 98-112 |
| Average (%) | 100 | 93 | 105 | |
| 1:4 | Range (%) | 94-110 | 82-96 | 99-115 |
| Average (%) | 102 | 88 | 105 | |
| 1:8 | Range (%) | 93-110 | 86-100 | 93-111 |
| Average (%) | 101 | 93 | 101 | |
| 1:16 | Range (%) | 95-110 | 83-98 | 101-115 |
| Average (%) | 103 | 90 | 108 |
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: Procyanidin B2 ameliorates free fatty acids-induced hepatic steatosis through regulating TFEB-mediated lysosomal pathway and redox state
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样品类型: Serum - 文献: Bifidobacterium adolescentis Isolated from Different Hosts Modifies the Intestinal Microbiota and Displays Differential Metabolic and Immunomodulatory Properties in Mice Fed a High-Fat Diet
样品类型: Serum - 文献: Asprosin, a novel glucogenic adipokine: a potential therapeutic implication in diabetes mellitus
样品类型: Serum - 文献: Hypothermic Effect of Acute Citral Treatment during LPS-induced Systemic Inflammation in Obese Mice: Reduction of Serum TNF-α and Leptin Levels
样品类型: Serum - 文献: A multi-targeting strategy to ameliorate high-fat-diet- and fructose-induced (western diet-induced) non-alcoholic fatty liver disease (NAFLD) with supplementation of a mixture of legume ethanol extracts
样品类型: Serum - 文献: B-Cell-Activating Factor Depletion Ameliorates Aging-Dependent Insulin Resistance via Enhancement of Thermogenesis in Adipose Tissues
样品类型: serum - 文献: The protective effects of Chinese yam polysaccharide against obesity-induced insulin resistance
样品类型: Serum - 文献: Pancreastatin inhibitor PSTi8 protects the obesity associated skeletal muscle insulin resistance in diet induced streptozotocin-treated diabetic mice
样品类型: plasma - 文献: Role of TRPV1/TRPV3 channels in olanzapine-induced metabolic alteration: Possible involvement in hypothalamic energy-sensing, appetite regulation, inflammation and mesolimbic pathway
样品类型: Serum - 文献: Modeling of antipsychotic-induced metabolic alterations in mice: An experimental approach precluding psychosis as a predisposing factor
样品类型: Serum - 文献: Berberine attenuated olanzapine-induced metabolic alterations in mice: Targeting transient receptor potential vanilloid type 1 and 3 channels
样品类型: serum - 文献: Virgin Grape Seed Oil Alleviates Insulin Resistance and Energy Metabolism Disorder in Mice Fed a High‐Fat Diet
样品类型: Serum - 文献: Astragalin, (3-O-glucoside of kaempferol), isolated from Moringa oleifera leaves modulates leptin, adiponectin secretion and inhibits adipogenesis in 3T3-L1 adipocytes
样品类型: 3T3-L1 cell - 文献: Antioxidant Activity of Sprouts Extracts Is Correlated with Their Anti-Obesity and Anti-Inflammatory Effects in High-Fat Diet-Fed Mice
样品类型: Plasma - 文献: Is the FVB/N mouse strain truly resistant to diet-induced obesity?
样品类型: serum
