Human HIF-1α(Hypoxia Inducible Factor 1 Alpha) ELISA Kit
人低氧诱导因子1α(HIF-1α)酶联免疫吸附测定试剂盒
规格: | / 96T / 48T / 96T*5 |
价格: | / ¥2800 / ¥1960 / ¥11900 |
货号:E-EL-H6066
反应性: 人
检测范围: 62.50~4000 pg/mL
灵敏度: 37.50 pg/mL
用途
该试剂盒用于体外定量检测人serum, plasma and other biological fluids中HIF-1α浓度
检测原理
本试剂盒采用双抗体夹心ELISA法。用抗人HIF-1α抗体包被于酶标板上,实验时样品或标准品中的人HIF-1α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HIF-1α抗体和辣根过氧化物酶标记的亲和素。抗人HIF-1α抗体与结合在包被抗体上的人HIF-1α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,HIF-1α浓度与OD450值之间呈正比,通过绘制标准曲线计算出样品中HIF-1α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 | 62.50—4000 pg/mL |
灵敏度 | 37.50 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HIF-1α,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 |
---|---|---|---|
4000 | 2.532 2.583 |
2.558 | 2.499 |
2000 | 1.676 1.61 |
1.643 | 1.584 |
1000 | 0.922 0.956 |
0.937 | 0.88 |
500 | 0.469 0.481 |
0.475 | 0.416 |
250 | 0.278 0.28 |
0.279 | 0.22 |
125 | 0.174 0.175 |
0.175 | 0.116 |
62.50 | 0.12 0.127 |
0.124 | 0.065 |
0 | 0.059 0.059 |
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 (pg/mL) |
177.29 | 422.96 | 1738.08 | 194.33 | 486.58 | 1980.59 |
Standard
deviation |
9.41 | 20.26 | 71.09 | 10.47 | 24.09 | 91.11 |
C V (%) | 5.31 | 4.79 | 4.09 | 5.39 | 4.95 | 4.60 |
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) | 94-109 | 102 |
EDTA plasma (n=5) | 99-109 | 104 |
Cell culture media (n=5) | 101-112 | 105 |
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 (%) | 88-101 | 94-106 | 91-102 |
Average (%) | 94 | 101 | 98 | |
1:4 | Range (%) | 93-107 | 94-104 | 92-105 |
Average (%) | 99 | 100 | 100 | |
1:8 | Range (%) | 86-98 | 90-100 | 96-110 |
Average (%) | 92 | 94 | 102 | |
1:16 | Range (%) | 94-108 | 99-107 | 84-99 |
Average (%) | 101 | 102 | 91 |
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: Comparison of Serum VEGF, IGF-1, and HIF-1α Levels in Children with Autism Spectrum Disorder and Healthy Controls
样品类型: Serum - 文献: Red blood cell transfusion induces abnormal HIF-1α response to cytokine storm after adult cardiac surgery
样品类型: serum - 文献: Semaphorin 3A attenuates the hypoxia suppression of osteogenesis in periodontal ligament stem cells
样品类型: Periodontal ligament stem cell (PDLSCs) - 文献: Cardiac rehabilitation may influence leptin and VEGF A crosstalk in patients after acute coronary syndrome
样品类型: serum - 文献: Shielding Engineered Islets With Mesenchymal Stem Cells Enhance Survival Under Hypoxia
样品类型: Cell lysate - 文献: Hypoxia-inducible factor-1α in acute ischemic stroke: neuroprotection for better clinical outcome
样品类型: Serum - 文献: Advanced platelet-rich fibrin promotes the paracrine function and proliferation of adipose-derived stem cells and contributes to micro-autologous fat transplantation by modulating HIF-1α and VEGF
样品类型: adipose-derived stem cell (ASCs) - 文献: Enhanced lysosome escape mediated by 1,2-dicarboxylic-cyclohexene anhydride-modified poly-l-lysine dendrimer as a gene delivery system
样品类型: Cell lysate - 文献: The administration of argon during ex vivo normothermic perfusion in an experimental model of kidney ischemia–reperfusion injury
样品类型: urine - 文献: The lower expression of circulating miR-210 and elevated serum levels of HIF-1α in ischemic stroke; Possible markers for diagnosis and disease prediction
样品类型: serum - 文献: Association between serum NLRP3 and malignant brain edema in patients with acute ischemic stroke
样品类型: serum - 文献: Role of TXNDC5 in tumorigenesis of colorectal cancer cells: In vivo and in vitro evidence
样品类型: Tissue homogenate - 文献: Hypoxia primed placental mesenchymal stem cells for wound healing
样品类型: Cell lysate - 文献: Expression and predictive value of HIF‑1α and VEGF in patients with burns following treatment
样品类型: serum - 文献: Expression and predictive value of HIF‑1α and VEGF in patients with burns following treatment
样品类型: Serum - 文献: Expression of early angiogenesis indicators in mature versus immature teeth
样品类型: Tissue homogenate - 文献: Based on HIF-1α/Wnt/β-Catenin Pathway to Explore the Effect of Qingshen Granules on Chronic Renal Failure Patients: A Randomized Controlled Trial
样品类型: Serum - 文献: HIF-1α Levels in patients receiving chemoradiotherapy for locally advanced non-small cell lung carcinoma
样品类型: serum - 文献: Hypoxia-induced inflammation and purinergic signaling in cross clamping the human aorta
样品类型: Serum