Rat KIM-1(Kidney Injury Molecule 1) ELISA Kit
大鼠肾损伤分子1(KIM-1)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-R3019
反应性: 大鼠
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大鼠serum, plasma and other biological fluids中KIM-1浓度
检测原理
本试剂盒采用双抗体夹心ELISA法。用抗大鼠KIM-1抗体包被于酶标板上，实验时样品或标准品中的大鼠KIM-1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大鼠KIM-1抗体和辣根过氧化物酶标记的亲和素。抗大鼠KIM-1抗体与结合在包被抗体上的大鼠KIM-1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，KIM-1浓度与OD450值之间呈正比，通过绘制标准曲线计算出样品中KIM-1的浓度。
| 反应类型 | 夹心法 |
| 规格 | 96T |
| 反应时间 | 3.5h |
| 反应性 | 大鼠 |
| 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
可检测样本中的大鼠KIM-1，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 | 2.448 2.597 |
2.523 | 2.464 |
| 10 | 1.68 1.644 |
1.662 | 1.603 |
| 5 | 0.963 0.928 |
0.946 | 0.887 |
| 2.5 | 0.495 0.48 |
0.488 | 0.429 |
| 1.25 | 0.266 0.277 |
0.272 | 0.213 |
| 0.63 | 0.184 0.186 |
0.185 | 0.126 |
| 0.31 | 0.12 0.116 |
0.118 | 0.059 |
| 0 | 0.056 0.061 |
0.059 | -- |
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) |
0.92 | 2.30 | 8.98 | 0.97 | 2.38 | 9.60 |
| Standard
deviation |
0.05 | 0.11 | 0.49 | 0.05 | 0.11 | 0.45 |
| C V (%) | 5.43 | 4.78 | 5.46 | 5.15 | 4.62 | 4.69 |
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) | 87-99 | 92 |
| EDTA plasma (n=5) | 87-101 | 95 |
| Cell culture media (n=5) | 90-101 | 94 |
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 (%) | 90-102 | 98-107 | 91-101 |
| Average (%) | 96 | 103 | 94 | |
| 1:4 | Range (%) | 90-98 | 87-97 | 92-100 |
| Average (%) | 95 | 90 | 96 | |
| 1:8 | Range (%) | 98-106 | 89-101 | 85-98 |
| Average (%) | 103 | 93 | 91 | |
| 1:16 | Range (%) | 94-108 | 95-108 | 90-101 |
| Average (%) | 101 | 103 | 96 |
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: Ameliorative Effects of a Rhenium (V) Compound with Uracil-Derived Ligand Markers Associated with Hyperglycaemia-Induced Renal Dysfunction in Diet-Induced Prediabetic Rats
样品类型: plasma - 文献: Renoprotective and therapeutic effects of newly water, ethanol, and butanol ginseng fractions in hypertensive and chronic kidney disease with L-NAME
样品类型: serum - 文献: Preventing the onset of diabetes-induced chronic kidney disease during prediabetes: The effects of oleanolic acid on selected markers of chronic kidney disease in a diet-induced prediabetic rat model
样品类型: Plasma - 文献: Epigenetic restoration of endogenous Klotho expression alleviates acute kidney injury-diabetes comorbidity
样品类型: urinary - 文献: Angiotensin 1–7 mitigates rhabdomyolysis induced renal injury in rats via modulation of TLR-4/NF-kB/iNOS and Nrf-2/heme‑oxygenase-1 signaling pathways
样品类型: serum - 文献: Agomelatine prevents gentamicin nephrotoxicity by attenuating oxidative stress and TLR-4 signaling, and upregulating PPARγ and SIRT1
样品类型: Serum - 文献: Ameliorative Effects of Bredemolic Acid on Markers Associated with Renal Dysfunction in a Diet-Induced Prediabetic Rat Model
样品类型: Plasma,Urine,Tissue homogenate - 文献: Pre‐ischemic renal lavage protects against renal ischemia‐reperfusion injury by attenuation of local and systemic inflammatory responses
样品类型: Urine - 文献: Piperazine ferulate attenuates gentamicin-induced acute kidney injury via the NF-κB/NLRP3 pathway
样品类型: serum - 文献: Saroglitazar, a dual PPAR-α/γ agonist, alleviates LPS-induced hepatic and renal injury in rats
样品类型: liver - 文献: Protective effect of fatty acid amide hydrolase inhibitor URB597 and monoacylglycerol lipase inhibitor KML29 on renal ischemia–reperfusion injury via toll-like receptor 4/nuclear factor-kappa B pathway
样品类型: serum - 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: Umbelliferone attenuates gentamicin-induced renal toxicity by suppression of TLR-4/NF-κB-p65/NLRP-3 and JAK1/STAT-3 signaling pathways
样品类型: Serum - 文献: Aged garlic extract rescues ethephon-induced kidney damage by modulating oxidative stress, apoptosis, inflammation, and histopathological changes in rats
样品类型: Serum - 文献: An in vivo and in vitro model on the protective effect of cilnidipine on contrast-induced nephropathy via regulation of apoptosis and CaMKⅡ/mPTP pathway
样品类型: serum - 文献: Candesartan Protects Against Cadmium-Induced Hepatorenal Syndrome by Affecting Nrf2, NF-κB, Bax/Bcl-2/Cyt-C, and Ang II/Ang 1–7 Signals
样品类型: kidney - 文献: Acetaminophen-Induced Nephrotoxicity: Suppression of Apoptosis and Endoplasmic Reticulum Stress Using Boric Acid
样品类型: serum - 文献: Effect of Hydrogen Sulfide on Kidney Injury in Rat Model of Crush Syndrome
样品类型: Tissue homogenate - 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: The protective effect of cannabinoid type 2 receptor activation on renal ischemia–reperfusion injury
样品类型: Serum - 文献: Protective effect of N‐(p‐amylcinnamoyl) anthranilic acid, phospholipase A2 enzyme inhibitor, and transient receptor potential melastatin‐2 channel blocker against renal ischemia–reperfusion injury
样品类型: Serum - 文献: Effects of hydrogen sulfide on acetaminophen-induced acute renal toxicity in rats
样品类型: Tissue homogenate - 文献: Beneficial effects of bardoxolone methyl, an Nrf2 activator, on crush-related acute kidney injury in rats
样品类型: Tissue homogenate
