Rat INS(Insulin) ELISA Kit
大鼠胰岛素(INS)酶联免疫吸附测定试剂盒
| 规格： | / 96T / 48T / 96T*5 |
| 价格： | / ￥2400 / ￥1650 / ￥10200 |
货号：E-EL-R2466
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cell culture supernatant and tissue homogenate中INS浓度
检测原理
本试剂盒采用双抗体夹心ELISA法。用抗大鼠INS抗体包被于酶标板上，实验时样品或标准品中的大鼠INS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INS抗体和辣根过氧化物酶标记的亲和素。抗大鼠INS抗体与结合在包被抗体上的大鼠INS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，INS浓度与OD450值之间呈正比，通过绘制标准曲线计算出样品中INS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 | cell culture supernatant and tissue homogenate |
特异性
可检测样本中的大鼠INS，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 | 2.415 2.451 |
2.433 | 2.38 |
| 10 | 1.53 1.588 |
1.559 | 1.506 |
| 5 | 0.888 0.87 |
0.879 | 0.826 |
| 2.5 | 0.433 0.439 |
0.436 | 0.383 |
| 1.25 | 0.208 0.196 |
0.202 | 0.149 |
| 0.63 | 0.153 0.143 |
0.148 | 0.095 |
| 0.31 | 0.097 0.107 |
0.102 | 0.049 |
| 0 | 0.047 0.059 |
0.053 | -- |
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) |
0.99 | 2.68 | 6.73 | 1.00 | 2.82 | 6.90 |
| Standard
deviation |
0.06 | 0.14 | 0.25 | 0.07 | 0.16 | 0.35 |
| C V (%) | 6.06 | 5.22 | 3.71 | 7.00 | 5.67 | 5.07 |
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) | ||
| EDTA plasma (n=5) | ||
| Cell culture media (n=5) | 86-98 | 92 |
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 (%) | 96-112 | ||
| Average (%) | 102 | |||
| 1:4 | Range (%) | 90-101 | ||
| Average (%) | 95 | |||
| 1:8 | Range (%) | 84-96 | ||
| Average (%) | 89 | |||
| 1:16 | Range (%) | 86-98 | ||
| Average (%) | 92 |
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: Potential mechanistic pathways underlying intestinal and hepatic effects of kefir in high-fructose-fed rats
样品类型: Tissue homogenate - 文献: GLP-1-Induced AMPK Activation Inhibits PARP-1 and Promotes LXR-Mediated ABCA1 Expression to Protect Pancreatic β-Cells Against Cholesterol-Induced Toxicity Through Cholesterol Efflux
样品类型: INS-1 cells - 文献: Replication Activities of Major 5′ Terminally Deleted Group-B Coxsackievirus RNA Forms Decrease PCSK2 mRNA Expression Impairing Insulin Maturation in Pancreatic Beta Cells
样品类型: INS-1 cells - 文献: Role of protein tyrosine phosphatase 1B inhibitor in central insulin resistance and associated cognitive deficits
样品类型: Tissue homogenate - 文献: Heme Oxygenase-1 (HMOX-1) and inhibitor of differentiation proteins (ID1, ID3) are key response mechanisms against iron-overload in pancreatic β-cells
样品类型: Pancreatic β cell - 文献: Sox9 is required in regeneration of pancreatic β cells following injury
样品类型: RIN-m5F cell - 文献: Bee bread mitigates downregulation of steroidogenic genes, decreased spermatogenesis, and epididymal oxidative stress in male rats fed with high-fat diet
样品类型: serum,testis - 文献: MKP-5 Relieves Lipotoxicity-Induced Islet β-Cell Dysfunction and Apoptosis via Regulation of Autophagy
样品类型: Cell lysate - 文献: Methylation and Acetylation Enhanced the Antidiabetic Activity of Some Selected Flavonoids: In Vitro, Molecular Modelling and Structure Activity Relationship-Based Study
样品类型: Cell culture medium - 文献: Modified Si-Miao-San ameliorates pancreatic B cell dysfunction by inhibition of reactive oxygen species-associated inflammation through AMP-kinase activation
样品类型: Cell culture supernatant - 文献: The Ameliorative Effect of Pectin-Insulin Patch On Renal Injury in Streptozotocin-Induced Diabetic Rats
样品类型: Urine,Tissue homogenate,Plasma - 文献: Nitric oxide interacts with cholinoceptors to modulate insulin secretion by pancreatic β cells
样品类型: Cell culture supernatant - 文献: Bone Marrow-Derived Mesenchymal Stem Cells Restored High-Fat-Fed Induced Hyperinsulinemia in Rats at Early Stage of Type 2 Diabetes Mellitus:
样品类型: Serum,plasma - 文献: Effects of noopept on cognitive functions and pubertal process in rats with diabetes
样品类型: Serum,Tissue homogenate - 文献: Orphan G-protein coupled receptor 183 (GPR183) potentiates insulin secretion and prevents glucotoxicity-induced β-cell dysfunction
样品类型: INS-1 (832/13) cells - 文献: The Chinese medicine Chai Hu Li Zhong Tang protects against non-alcoholic fatty liver disease by activating AMPKα
样品类型: Cell culture supernatant - 文献: Let7b-5p is Upregulated in the Serum of Emirati Patients with Type 2 Diabetes and Regulates Insulin Secretion in INS-1 Cells
样品类型: INS-1 (832/13) cell - 文献: Protective and Therapeutic Effects of Orlistat on Metabolic Syndrome and Oxidative Stress in High-Fat Diet-Induced Metabolic Dysfunction-Associated Fatty Liver Disease (MAFLD) in Rats: Role on Nrf2 Activation
- 文献: Cotransplantation of Mesenchymal Stem Cells and Immature Dendritic Cells Potentiates the Blood Glucose Control of Islet Allografts
样品类型: Cell culture supernatant - 文献: GNAS gene is an important regulator of insulin secretory capacity in pancreatic β-cells
样品类型: INS -1 (832/13) cells - 文献: Protective effects of essential oil from Fructus Alpiniae zerumbet on retinal Müller gliosis via the PPAR-γ-p-CREB signaling pathway
样品类型: Serum,Cell culture medium - 文献: Aqueous leaf extract of Clinacanthus nutans improved metabolic indices and sorbitol‐related complications in type II diabetic rats (T2D)
样品类型: Tissue homogenate,Serum - 文献: CGA ameliorates cognitive decline by regulating the PI3K/AKT signaling pathway and neurotransmitter systems in rats with multi-infarct dementia
样品类型: Tissue homogenate - 文献: RORB and RORC associate with human islet dysfunction and inhibit insulin secretion in INS-1 cells
样品类型: INS-1 (832/13) cells - 文献: Reduced Expression of Chl1 gene Impairs Insulin Secretion by Down-Regulating the Expression of Key Molecules of β-cell Function
样品类型: Cell culture supernatant
