ALD(Aldosterone) ELISA Kit
醛固酮(ALD)酶联免疫吸附测定试剂盒
| 规格： | / 96T / 48T / 96T*5 |
| 价格： | / ￥2600 / ￥1800 / ￥11050 |
货号：E-EL-0070
反应性: 通用
检测范围: 31.25~2000 pg/mL
灵敏度: 18.75 pg/mL
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ALD浓度。
检测原理
本试剂盒采用竞争ELISA法。用ALD抗原包被于酶标板上，实验时样品或标准品中的ALD与包被的ALD竞争生物素标记的抗ALD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，ALD浓度与OD450值之间呈反比，通过绘制标准曲线计算出样品中ALD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ALD，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 | 0.306 0.354 |
0.33 |
| 1000 | 0.42 0.434 |
0.427 |
| 500 | 0.612 0.594 |
0.603 |
| 250 | 0.87 0.898 |
0.884 |
| 125 | 1.259 1.253 |
1.256 |
| 62.5 | 1.647 1.635 |
1.641 |
| 31.25 | 1.949 1.951 |
1.95 |
| 0 | 2.382 2.39 |
2.386 |
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) |
51.80 | 89.60 | 476.90 | 46.80 | 94.20 | 479.40 |
| Standard
deviation |
2.70 | 4.30 | 20.00 | 3.20 | 5.70 | 23.00 |
| C V (%) | 5.21 | 4.80 | 4.19 | 6.84 | 6.05 | 4.80 |
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) | 86-98 | 91 |
| EDTA plasma (n=5) | 96-112 | 102 |
| Cell culture media (n=5) | 90-102 | 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 (%) | 91-106 | 88-101 | |
| Average (%) | 105 | 98 | 95 | |
| 1:4 | Range (%) | 94-107 | 87-103 | 87-98 |
| Average (%) | 101 | 95 | 92 | |
| 1:8 | Range (%) | 101-117 | 95-106 | 85-98 |
| Average (%) | 107 | 100 | 91 | |
| 1:16 | Range (%) | 97-109 | 87-101 | 89-99 |
| Average (%) | 104 | 94 | 94 | |
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
- 文献: Maternal High-Fructose Intake Induces Multigenerational Activation of the Renin-Angiotensin-Aldosterone System
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样品类型: Serum,urine - 文献: Caveolin 1-related autophagy initiated by aldosterone-induced oxidation promotes liver sinusoidal endothelial cells defenestration
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样品类型: Tissue homogenate - 文献: Amelioration of risk factors associated with diabetic nephropathy in diet-induced pre-diabetic rats by an uracil-derived diimine ruthenium(II) compound
样品类型: Plasma,Tissue homogenate - 文献: var kunstleri in spontaneously hypertensive rats: possible involvement of renin–angiotensin–aldosterone system, endothelial function and anti-oxidant system
样品类型: serum,urine - 文献: Oral hormonal therapy with ethinylestradiol–levonorgestrel improves insulin resistance, obesity, and glycogen synthase kinase-3 independent of circulating mineralocorticoid in estrogen-deficient rats
样品类型: plasma - 文献: Ameliorative effect of low-dose spironolactone on obesity and insulin resistance is through replenishment of estrogen in ovariectomized rats
样品类型: Plasma - 文献: Anti-hyperglycaemic effects of dioxidovanadium complex cis-[VO2(obz)py] avert kidney dysfunction in streptozotocin-induced diabetic male Sprague–Dawley rats
样品类型: plasma - 文献: Long-term exposure to crotonaldehyde causes heart and kidney dysfunction through induction of inflammatory and oxidative damage in male Wistar rats
样品类型: heart tissue - 文献: Aliskiren, Fosinopril, and Their Outcome on Renin-Angiotensin-Aldosterone System (RAAS) in Rats with Thyroid Dysfunction
样品类型: serum - 文献: The effect of eplerenone on the renin‐angiotensin‐aldosterone system of rats with thyroid dysfunction
样品类型: Serum - 文献: Cardio-protective effects of a dioxidovanadium(V) complex in male sprague–dawley rats with streptozotocin-induced diabetes
样品类型: Plasma - 文献: Allopurinol and valproic acid improve cardiac triglyceride and Na+-K+-ATPase activity independent of circulating aldosterone in female rats with glucose intolerance
样品类型: plasma - 文献: Neuroglobin correlates with cryptochrome-1 in obstructive sleep apnea with primary aldosteronism
样品类型: Serum - 文献: Two Sulfur Glycoside Compounds Isolated from Lepidium apetalum Willd Protect NRK52e Cells against Hypertonic-Induced Adhesion and Inflammation by Suppressing the MAPK Signaling Pathway and RAAS
样品类型: Cell culture supernatant - 文献: The Effects of Momordica balsamina Methanolic Extract on Kidney Function in STZ-Induced Diabetic Rats: Effects on Selected Metabolic Markers
样品类型: plasma - 文献: Histone deacetylase inhibitor, mocetinostat, regulates cardiac remodelling and renin-angiotensin system activity in rats with transverse aortic constriction-induced pressure overload cardiac hypertrophy.
样品类型: serum - 文献: Nicotine exposure suppresses hyperinsulinemia and improves endothelial dysfunction mediators independent of corticosteroids in insulin-resistant oral contraceptive-treated female rats
样品类型: Plasma - 文献: Maternal high-fructose intake during pregnancy and lactation induces metabolic syndrome in adult offspring
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样品类型: Plasma,Serum,Tissue homogenate
