Human KL(Klotho) ELISA Kit
人可罗索(KL)酶联免疫吸附测定试剂盒
| 规格： | / 96T / 48T / 96T*5 |
| 价格： | / ￥3200 / ￥2300 / ￥13600 |
货号：E-EL-H5451
反应性: 人
检测范围: 0.31~20 ng/mL
灵敏度: 0.19 ng/mL
用途
该试剂盒用于体外定量检测人serum, plasma and other biological fluids中KL浓度
检测原理
本试剂盒采用双抗体夹心ELISA法。用抗人KL抗体包被于酶标板上，实验时样品或标准品中的人KL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KL抗体和辣根过氧化物酶标记的亲和素。抗人KL抗体与结合在包被抗体上的人KL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，KL浓度与OD450值之间呈正比，通过绘制标准曲线计算出样品中KL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 | 0.31—20 ng/mL |
| 灵敏度 | 0.19 ng/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KL，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 |
|---|---|---|---|
| 20 | 2.219 2.253 |
2.236 | 2.16 |
| 10 | 1.434 1.444 |
1.439 | 1.363 |
| 5 | 0.871 0.847 |
0.859 | 0.783 |
| 2.5 | 0.444 0.456 |
0.45 | 0.374 |
| 1.25 | 0.27 0.242 |
0.256 | 0.18 |
| 0.63 | 0.177 0.155 |
0.166 | 0.09 |
| 0.31 | 0.112 0.132 |
0.122 | 0.046 |
| 0 | 0.070 0.082 |
0.076 | -- |
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) |
1.04 | 2.05 | 9.74 | 0.97 | 2.04 | 9.08 |
| Standard
deviation |
0.06 | 0.11 | 0.34 | 0.05 | 0.10 | 0.48 |
| C V (%) | 5.77 | 5.37 | 3.49 | 5.15 | 4.90 | 5.29 |
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) | 86-99 | 91 |
| EDTA plasma (n=5) | 89-102 | 96 |
| Cell culture media (n=5) | 85-98 | 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 (%) | 99-114 | 97-113 | 97-108 |
| Average (%) | 106 | 105 | 102 | |
| 1:4 | Range (%) | 89-101 | 81-92 | 81-93 |
| Average (%) | 95 | 87 | 87 | |
| 1:8 | Range (%) | 89-100 | 87-102 | 85-100 |
| Average (%) | 95 | 93 | 91 | |
| 1:16 | Range (%) | 94-107 | 80-90 | 88-102 |
| Average (%) | 99 | 85 | 93 |
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: AB0240 CORRELATION BETWEEN KLOTHO LEVELS AND ANKLE-BRACHIAL INDEX IN FEMALE RHEUMATOID ARTHRITIS PATIENTS
样品类型: serum - 文献: Efficacy of a novel proprietary dietary supplement (TRI 360TM) on psychological symptoms and stress-related quality of life in adult subjects: A randomized controlled clinical trial
样品类型: serum - 文献: CCL5 Suppresses Klotho Expression via p-STAT3/DNA Methyltransferase1-Mediated Promoter Hypermethylation
样品类型: Serum - 文献: Kernelized k-Local Hyperplane Distance Nearest-Neighbor Model for Predicting Cerebrovascular Disease in Patients With End-Stage Renal Disease
样品类型: serum - 文献: Alpha-Klotho protein in systemic lupus erythematosus
样品类型: Serum - 文献: The Association of Fibroblast Growth Factor 23 with Arterial Stiffness and Atherosclerosis in Patients with Autosomal Dominant Polycystic Kidney Disease
样品类型: serum - 文献: Enhancement of Apo2L/TRAIL signaling pathway receptors by the activation of Klotho gene with CRISPR/Cas9 in Caco-2 colon cancer cells
样品类型: Caco-2 cell - 文献: Serum Klotho as a marker for early diagnosis of acute kidney injury after cardiac surgery
样品类型: Serum - 文献: A Comprehensive Study on the Anti-cancer Effects of Quercetin and Its Epigenetic Modifications in Arresting Progression of Colon Cancer Cell Proliferation
样品类型: L132 cell,HCT 116 cell,COLO 320 cell,COLO 205 cell - 文献: Body composition, adipokines, FGF23-Klotho and bone in kidney transplantation: Is there a link?
样品类型: Serum - 文献: Evaluation of serum and aqueous humor klotho levels in pseudoexfoliation syndrome, pseudoexfoliation and primary open-angle glaucoma
样品类型: serum - 文献: Klotho, vitamin D and homocysteine levels during acute episode and remission periods in schizophrenia patients
样品类型: Serum - 文献: A Self-Representation-Based Fuzzy SVM Model for Predicting Vascular Calcification of Hemodialysis Patients
样品类型: serum - 文献: Association of plasma neutrophil gelatinase-associated lipocalin and thoracic aorta calcification in maintenance hemodialysis patients with and without diabetes
样品类型: plasma - 文献: Association of plasma neutrophil gelatinase-associated lipocalin with parameters of CKD–MBD in maintenance hemodialysis patients
样品类型: plasma - 文献: The correlation between fibroblast growth factor-23 and ESRD patients with hearing impairment
样品类型: serum - 文献: Association of serum FGF-23, klotho, fetuin-A, osteopontin, osteoprotegerin and hs-CRP levels with coronary artery disease
样品类型: serum - 文献: Comparison of serum human Klotho levels and thiol/disulfide homeostasis in women with polycystic ovary syndrome and in healthy women
样品类型: Serum - 文献: The role of biofield energy treatment on psychological symptoms, mental health disorders, and stress-related quality of life in adult subjects: A randomized controlled clinical trial
样品类型: serum
