Human MMP-13(Matrix Metalloproteinase 13) ELISA Kit
人基质金属蛋白酶13(MMP-13)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-H6023
反应性: 人
检测范围: 15.63~1000 pg/mL
灵敏度: 9.38 pg/mL
用途
该试剂盒用于体外定量检测人serum, plasma and other biological fluids中MMP-13浓度
检测原理
本试剂盒采用双抗体夹心ELISA法。用抗人MMP-13抗体包被于酶标板上，实验时样品或标准品中的人MMP-13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MMP-13抗体和辣根过氧化物酶标记的亲和素。抗人MMP-13抗体与结合在包被抗体上的人MMP-13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-13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-13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 | 15.63—1000 pg/mL |
| 灵敏度 | 9.38 pg/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MMP-13，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 | Corrected |
|---|---|---|---|
| 1000 | 2.564 2.571 |
2.568 | 2.507 |
| 500 | 1.674 1.645 |
1.66 | 1.599 |
| 250 | 0.907 0.97 |
0.937 | 0.878 |
| 125 | 0.463 0.458 |
0.461 | 0.4 |
| 62.5 | 0.28 0.271 |
0.276 | 0.215 |
| 31.25 | 0.172 0.172 |
0.172 | 0.111 |
| 15.63 | 0.112 0.121 |
0.117 | 0.056 |
| 0 | 0.061 0.06 |
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 (pg/mL) |
43.41 | 107.90 | 450.13 | 44.04 | 101.12 | 413.83 |
| Standard
deviation |
2.34 | 4.95 | 22.82 | 2.53 | 5.06 | 19.66 |
| C V (%) | 5.39 | 4.59 | 5.07 | 5.74 | 5.00 | 4.75 |
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) | 95-106 | 98 |
| EDTA plasma (n=5) | 98-110 | 105 |
| Cell culture media (n=5) | 90-103 | 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 (%) | 95-109 | 86-96 | 90-101 |
| Average (%) | 101 | 91 | 97 | |
| 1:4 | Range (%) | 84-95 | 92-103 | 98-106 |
| Average (%) | 91 | 96 | 102 | |
| 1:8 | Range (%) | 97-104 | 92-105 | 89-102 |
| Average (%) | 101 | 100 | 96 | |
| 1:16 | Range (%) | 87-101 | 96-110 | 93-99 |
| Average (%) | 94 | 104 | 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: Associations between serum S100A8/S100A9 and knee symptoms, joint structures and cartilage enzymes in patients with knee osteoarthritis
样品类型: Serum - 文献: Single Cell RNA-Seq Analysis Identifies Differentially Expressed Genes of Treg Cell in Early Treatment-Naive Rheumatoid Arthritis By Arsenic Trioxide
样品类型: Cell sample - 文献: Extracellular vesicle and soluble fractions of adipose tissue-derived mesenchymal stem cells secretome induce inflammatory cytokines modulation in an in vitro model of discogenic pain
样品类型: human adipose-derived mesenchymal stem cells (HAdMSC),h - 文献: Associations between serum ghrelin and knee symptoms, joint structures and cartilage or bone biomarkers in patients with knee osteoarthritis
样品类型: Serum - 文献: Arsenic trioxide improves Treg and Th17 balance by modulating STAT3 in treatment-naïve rheumatoid arthritis patients
样品类型: PBMCs cells - 文献: Arsenic Trioxide in Synergy with Vitamin D Rescues the Defective VDR-PPAR- Functional Module of Autophagy in Rheumatoid Arthritis
样品类型: rheumatoid arthritis (RA) fibroblast-like synoviocytes - 文献: Ferroptosis Plays a Role in Human Chondrocyte of Osteoarthritis Induced by IL-1β In Vitro
样品类型: Chondrocyte - 文献: The protective effects of Olmesartan against interleukin-29 (IL-29)-induced type 2 collagen degradation in human chondrocytes
样品类型: T/C-28a2 cell - 文献: MicroRNA-449a upregulation promotes chondrocyte extracellular matrix degradation in osteoarthritis
样品类型: Cell culture supernatant - 文献: Ethanolic extract of Kaempferia parviflora interrupts the mechanisms-associated rheumatoid arthritis in SW982 culture model via p38/STAT1 and STAT3 pathways
样品类型: Cell culture supernatant - 文献: MicroRNA-132 upregulation promotes matrix degradation in intervertebral disc degeneration
样品类型: Cell culture supernatant - 文献: Associations between serum IL-8 and knee symptoms, joint structures, and cartilage or bone biomarkers in patients with knee osteoarthritis
样品类型: Serum - 文献: Vicenin 3 ameliorates ECM degradation by regulating the MAPK pathway in SW1353 chondrocytes
样品类型: SW1353 cell - 文献: Association of IL-6, IL-8, MMP-13 gene polymorphisms with knee osteoarthritis susceptibility in the Chinese Han population
样品类型: Tissue homogenate - 文献: Potential Value of Matrix Metalloproteinase-13 as a Biomarker for Osteoarthritis
样品类型: serum - 文献: Protective effects of stem cells from human exfoliated deciduous teeth derived conditioned medium on osteoarthritic chondrocytes
样品类型: Cell culture medium - 文献: Circ-LRP1B functions as a competing endogenous RNA to regulate proliferation, apoptosis and oxidative stress of LPS-induced human C28/I2 chondrocytes
样品类型: C28/I2 cell - 文献: Expression and Significance of MMPs in Synovial Fluid, Serum and PBMC Culture Supernatant Stimulated by LPS in Osteoarthritis Patients With or Without Diabetes
样品类型: Serum,Synovia - 文献: Chrysin protects human osteoarthritis chondrocytes by inhibiting inflammatory mediator expression via HMGB1 suppression
样品类型: Cell culture medium - 文献: Associations of serum citrate levels with knee structural changes and cartilage enzymes in patients with knee osteoarthritis
样品类型: serum
