Rat MMP-9(Matrix Metalloproteinase 9) ELISA Kit
大鼠基质金属蛋白酶9(MMP-9)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-R3021
反应性: 大鼠
检测范围: 7.81~500 ng/mL
灵敏度: 4.69 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MMP-9浓度
检测原理
本试剂盒采用双抗体夹心ELISA法。用抗大鼠MMP-9抗体包被于酶标板上，实验时样品或标准品中的大鼠MMP-9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MMP-9抗体和辣根过氧化物酶标记的亲和素。抗大鼠MMP-9抗体与结合在包被抗体上的大鼠MMP-9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，MMP-9浓度与OD450值之间呈正比，通过绘制标准曲线计算出样品中MMP-9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 | 7.81—500 ng/mL |
| 灵敏度 | 4.69 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MMP-9，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 |
|---|---|---|---|
| 500 | 2.514 2.51 |
2.512 | 2.451 |
| 250 | 1.698 1.649 |
1.674 | 1.613 |
| 125 | 0.995 0.92 |
0.958 | 0.897 |
| 62.5 | 0.46 0.513 |
0.487 | 0.426 |
| 31.25 | 0.268 0.261 |
0.265 | 0.204 |
| 15.63 | 0.177 0.188 |
0.183 | 0.122 |
| 7.81 | 0.122 0.118 |
0.12 | 0.059 |
| 0 | 0.056 0.065 |
0.061 | -- |
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) |
23.64 | 61.14 | 223.70 | 24.02 | 56.48 | 214.26 |
| Standard
deviation |
1.41 | 2.78 | 9.91 | 1.21 | 2.80 | 11.48 |
| C V (%) | 5.96 | 4.55 | 4.43 | 5.04 | 4.96 | 5.36 |
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) | 98-104 | 101 |
| EDTA plasma (n=5) | 95-109 | 103 |
| Cell culture media (n=5) | 91-101 | 95 |
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 (%) | 86-95 | 89-97 | 92-103 |
| Average (%) | 90 | 94 | 96 | |
| 1:4 | Range (%) | 94-104 | 97-110 | 99-111 |
| Average (%) | 98 | 102 | 104 | |
| 1:8 | Range (%) | 98-105 | 90-98 | 87-99 |
| Average (%) | 102 | 95 | 93 | |
| 1:16 | Range (%) | 91-104 | 94-109 | 94-104 |
| Average (%) | 98 | 102 | 97 |
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: Protective Effect of Liposomal Epigallocatechin-Gallate in Experimental Gentamicin-Induced Hepatotoxicity
样品类型: serum - 文献: Water-soluble alkaloids extracted from Aconiti Radix lateralis praeparata protect against chronic heart failure in rats via a calcium signaling pathway
样品类型: Plasma - 文献: Huangbai liniment and berberine promoted wound healing in high-fat diet/Streptozotocin-induced diabetic rats
样品类型: skin tissue - 文献: Coomassie brilliant blue G-250 dye attenuates bleomycin-induced lung fibrosis by regulating the NF-κB and NLRP3 crosstalk: A novel approach for filling an unmet medical need
样品类型: lung - 文献: The Activation of ROS/NF-κB/MMP-9 Pathway Promotes Calcium-Induced Kidney Crystal Deposition
样品类型: serum - 文献: Intragastric administration of Pien Tze Huang enhanced wound healing in diabetes by inhibiting inflammation and improving energy generation
样品类型: skin tissue - 文献: Metabolites of gut microbiota fermenting Poria cocos polysaccharide alleviates chronic nonbacterial prostatitis in rats
样品类型: prostate - 文献: Fabrication of In Situ Layered Hydrogel Scaffold for the Co-delivery of PGDF-BB/Chlorhexidine to Regulate Proinflammatory Cytokines, Growth Factors, and MMP-9 in a Diabetic Skin Defect Albino Rat Model
样品类型: Tissue homogenate,Serum - 文献: Niacin mitigates blood–brain barrier tight junctional proteins dysregulation and cerebral inflammation in ketamine rat model of psychosis: Role of GPR109A receptor
样品类型: Brain tissue - 文献: Effect of chronic left ventricular unloading on myocardial remodeling: Multimodal assessment of two heterotopic heart transplantation techniques
样品类型: Tissue homogenate - 文献: The Mechanisms of Cucurbitacin E as a Neuroprotective and Memory-Enhancing Agent in a Cerebral Hypoperfusion Rat Model: Attenuation of Oxidative Stress, Inflammation, and Excitotoxicity.
样品类型: brain - 文献: l-Borneol Exerted the Neuroprotective Effect by Promoting Angiogenesis Coupled With Neurogenesis via Ang1-VEGF-BDNF Pathway
样品类型: Serum - 文献: HGF and TSG-6 Released by Mesenchymal Stem Cells Attenuate Colon Radiation-Induced Fibrosis
样品类型: Tissue homogenate - 文献: Network pharmacology analysis uncovers the effect on apoptotic pathway by Bu-Fei formula for COPD treatment
样品类型: serum - 文献: Integrated serum pharmacochemistry and network pharmacological analysis used to explore possible anti-rheumatoid arthritis mechanisms of the Shentong-Zhuyu decoction
样品类型: Serum - 文献: Exposure to Air Pollution Exacerbates Inflammation in Rats with Preexisting COPD
样品类型: Tissue homogenate,Bronchoalveolar Lavage Fluid - 文献: Nisin reduces uterine inflammation in rats by modulating concentrations of pro‐ and anti‐inflammatory cytokines
样品类型: Serum - 文献: Matrix metalloproteinase 9 a potential major player connecting atherosclerosis and osteoporosis in high fat diet fed rats
样品类型: Tissue homogenate - 文献: Inhibition effect of Caragana sinica root extracts on Osteoarthritis through MAPKs, NF-κB signaling pathway
样品类型: serum - 文献: Design, in vitro bioactivity and in vivo influence on oxidative stress and matrix metalloproteinases of bioglasses in experimental skin wound
样品类型: skin - 文献: St. Thomas and del Nido cardioplegia are superior to Custodiol cardioplegia in a rat model of donor heart
样品类型: Tissue homogenate - 文献: Neuroprotective Effects of Brain-Derived Neurotrophic Factor and Noggin-Modified Bone Mesenchymal Stem Cells in Focal Cerebral Ischemia in Rats
样品类型: Cell culture supernatant - 文献: The antiiflammatory effect of Ginkgo biloba in lipopolysaccharide-induced rat sepsis model
样品类型: Serum
