Human TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit

产品货号:E-EL-H0109c

规格: / 96T / 48T
价格: / ¥2400 / ¥1650

货号:E-EL-H0109c

货期:3个工作日+顺丰周期

反应性:

检测范围: 7.81~500 pg/mL

灵敏度: 4.69 pg/mL

  • 详情
  • 文献(32)
  • 用途

    该试剂盒用于体外定量检测人Serum, plasma and other biological fluids中天然及部分重组TNF-α浓度

    检测原理

    本试剂盒采用双抗体夹心ELISA法。用抗人TNF-α抗体包被于酶标板上,实验时样品或标准品中的人TNF-α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TNF-α抗体和辣根过氧化物酶标记的亲和素。抗人TNF-α抗体与结合在包被抗体上的人TNF-α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,TNF-α浓度与OD450值之间呈正比,通过绘制标准曲线计算出样品中TNF-α的浓度。

    反应类型 夹心法
    规格 96T
    反应时间 3.5h
    反应性
    检测方法 Colormetric
    检测范围 7.81—500 pg/mL
    灵敏度 4.69 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TNF-α,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
    500 2.427
    2.445
    2.436 2.376
    250 1.562
    1.584
    1.573 1.513
    125 0.993
    0.979
    0.986 0.926
    62.5 0.477
    0.515
    0.496 0.436
    31.25 0.245
    0.237
    0.241 0.181
    15.63 0.166
    0.16
    0.163 0.103
    7.81 0.105
    0.121
    0.113 0.053
    0 0.059
    0.061
    0.06 --

    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)
    24.10 79.50 183.00 22.70 86.50 177.40
    Standard
    deviation
    1.50 3.80 9.70 1.20 4.00 7.60
    C V (%) 6.22 4.78 5.30 5.29 4.62 4.28

    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) 86-100 92
    EDTA plasma (n=5) 94-106 99
    Cell culture media (n=5) 93-107 99

    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 (%) 92-105 84-97 88-99
    Average (%) 99 91 93
    1:4 Range (%) 99-116 87-99 91-104
    Average (%) 107 93 98
    1:8 Range (%) 97-113 81-93 92-109
    Average (%) 104 88 99
    1:16 Range (%) 96-112 85-98 95-107
    Average (%) 103 90 100

    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

    1. 文献: Jasim N A, Kamal Y M, Mohammed M M, et al. 14. Evaluation of the Anti-Inflammatory Effect of Telmisartan As an Adjuvant Therapy to NSAID in The Management of Knee Osteoarthritis; A Clinical Prospective Study[J]. Iraqi JMS 2018; Vol. 16(1): 100-110.
      样品类型: Serum
    2. 文献: Ruan G, Xu J, Wang K, et al. Associations between knee structural measures, circulating inflammatory factors and MMP13 in patients with knee osteoarthritis[J]. Osteoarthritis and cartilage, 2018.
      样品类型: Serum
    3. 文献: Saleh H H, Abdulridha M K, Najeb A A. Potential Benefit of Melatonin as an Adjuvant Therapy in a Sample of Iraqi Patients with Crohn’s Disease: A New Study[J]. ALMUSTANSIRIYAH Journal of PHARMACEUTICAL SCIENCES (AJPS), 2018, 17(2): 9-9.
      样品类型: Serum
    4. 文献: Zhou P, Liu H, Wu Y, et al. Propofol Promotes Ankle Fracture Healing in Children by Inhibiting Inflammatory Response[J]. Medical Science Monitor: International Medical Journal of Experimental And Clinical Research, 2018, 24: 4379.
      样品类型: Serum
    5. 文献: Wang F, Li X, Huang L, et al. High-Mobility Group Protein Box 1 is Upregulated in Children with Henoch-Schonlein Purpura[J]. Pediatric Allergy, Immunology, and Pulmonology, 2018, 31(2): 66-72.
      样品类型: Serum
    6. 文献: Akash M S H, Rehman K, Liaqat A, et al. Biochemical investigation of gender-specific association between insulin resistance and inflammatory biomarkers in types 2 diabetic patients[J]. Biomedicine & Pharmacotherapy, 2018, 106: 285-291.
      样品类型: Serum
    7. 文献: Giusti I, Di Francesco M D, D'Ascenzo S, et al. Leukocyte Depletion Does Not Affect The In Vitro Healing Ability Of Platelet Rich Plasma[J]. Experimental and Therapeutic Medicine, 2018, 15(4): 4029-4038.
    8. 文献: Hatef Z S, Nada S Z, Jasim A M. Adiponectin may be used as a marker in prediction of diabetic retinopathy patients[J]. Iraq Medical Journal, 2018, 2(1): 20-23.
      样品类型: Serum
    9. 文献: Wang C M, Zhang T, Wang Y H, et al. The Shortening Telomere Length Of T Lymphocytes Maybe Associated With HyperFunction In Servere Aplastic Anemia[J]. Molecular Medicine Reports, 2018, 17: 1015-1021.
    10. 文献: Lu A Q, Wang Z R, Wang S C. Role Of miR-589-3p In Human Lumbar Disc Degeneration And Its Potential Mechanism[J]. Experimental and Therapeutic Medicine, 2018, 15: 1616-1621.
      样品类型: Cell culture supernatant
    11. 文献: Pan Y Y, Zhou F H, Song Z H, et al. Oleanolic acid protects against pathogenesis of atherosclerosis, possibly via FXR-mediated angiotensin (Ang)-(1-7) upregulation[J]. Biomedicine & Pharmacotherapy, 2018, 97: 1694-1700.
      样品类型: Cell culture supernatant,Serum
    12. 文献: Al-Assadi N A M, Al-Qaysi S A. Tumor Necrosis Factor Alpha Synergetic with Insulin Resistance Potentially Contributes to the Development of Polycystic Ovary Syndrome[J]. Medical Journal of Babylon, 2017, 14(4): 732-737.
      样品类型: Serum
    13. 文献: Nada S Z, Hatef Z S, Alkutubi H M, et al. Serum Tnfα Levels Are Inversely Associated With Insulin Sensitivity in Diabetic Retinopathy Patients[J]. Kerbala Jorunal of Medicine, 2017, 10(3): 3857-3862.
      样品类型: Serum
    14. 文献: Montico B, Lapenta C, Ravo M, et al. Exploiting A New Strategy To Induce Immunogenic Cell Death To Improve Dendritic Cell-Based Vaccines For Lymphoma Immunotherapy[J]. OncoImmunology, 2017.
      样品类型: Cell culture supernatant
    15. 文献: Yu H, Dong R, Lu Y Y, et al. Short-Term Postoperative Cognitive Dysfunction And Inflammatory Response In Patients Undergoing Cytoreductive Surgery And Hyperthermic Intraperitoneal Chemotherapy: A Pilot Study[J]. Mediators Of Inflammation, 2017.
      样品类型: Serum
    16. 文献: Jiang Z, Wang W H, Guo C C. Tetrahydroxy Stilbene Glucoside Ameliorates H2O2-Induced Human Brain Microvascular Endothelial Cell Dysfunction In Vitro By Inhibiting Oxidative Stress And Inflammatory Responses[J]. Molecular Medicine Reports, 2017, 16(4): 5219-5224.
      样品类型: Cell culture supernatant
    17. 文献: Li Z, Tang Y, Tang N, et al. High Anti-Human Cytomegalovirus Antibody Levels Are Associated With The Progression Of Essential Hypertension And Target Organ Damage In Han Chinese Population[J]. Plos One, 2017.
      样品类型: Serum
    18. 文献: Cao Z, Chen L. Inhibition Of miR27a Suppresses The Inflammatory Response Via The p38/MAPK Pathway In Intervertebral Disc Cells[J]. Experimental and Therapeutic Medicine, 2017, 14(5): 4572-4578.
      样品类型: Cell culture supernatant
    19. 文献: Yan Z, Wang W, Wu Y, et al. Zinc Oxide Nanoparticle-Induced Atherosclerotic Alterations In Vitro And In Vivo[J]. International Journal of Nanomedicine, 2017, 12: 4433-4442.
      样品类型: Cell culture supernatant,Serum
    20. 文献: Belvedere R, Bizzarro V, Parente L, et al. The Pharmaceutical Device Prisma Skin Promotes In Vitro Angiogenesis Through Endothelial To Mesenchymal Transition During Skin Wound Healing[J]. International Journal of Molecular Sciences, 2017.
      样品类型: Cell culture supernatant
    21. 文献: Jiang Y, Kou J, Han X, et al. Ros-Dependent Activation Of Autophagy Through The PI3K/Akt/mTOR Pathway Is Induced By Hydroxysafflor Yellow A-Sonodynamic Therapy In THP-1 Macrophages[J]. Oxidative Medicine and Cellular Longevity, 2017.
      样品类型: Cell culture supernatant
    22. 文献: Chen W H, Luo G F, Lei Q, et al. Overcoming The Heat Endurance Of Tumor Cells By Interfering The Anaerobic Glycolysis Metabolism For Improved Photothermal Therapy[J]. Acs Nano, 2017, 11(2): 1419-1431.
      样品类型: Cell culture supernatant
    23. 文献: He W, Ren Y, Wang X, et al. C Reactive Protein And Enzymatically Modified LDL Cooperatively Promote Dendritic Cell-Mediated T Cell Activation[J]. Cardiovascular Pathology, 2017, 29: 1-6.
      样品类型: Cell culture supernatant
    24. 文献: Liu Q, Xu C, Ji G, et al. Sublethal Effects Of Zinc Oxide Nanoparticles On Male Reproductive Cells[J]. Toxicology in Vitro, 2016, 35: 131-138.
      样品类型: Cell culture supernatant
    25. 文献: Zhao Y, Jiang C, He J, et al. Multifunctional Dextran Sulfate-Coated Reconstituted High Density Lipoproteins Target Macrophages And Promote Beneficial Antiatherosclerotic Mechanisms[J]. Bioconjugate Chemistry, 2016.
      样品类型: Cell culture supernatant
    26. 文献: Zhang X P, Shang W L, Yuan J Z, et al. Positive Feedback Cycle Of TNF-α Promotes Staphylococcal Enterotoxin B-Induced THP-1 Cell Apoptosis[J]. Frontier in Cellular and Infection Microbiology, 2016.
      样品类型: Cell culture supernatant
    27. 文献: Cheng P, Hua W, Zhang W J, et al. Inhibitory Effect Of miR-125b On Hepatitis C Virus Core Protein-Induced TLR2/MyD88 Signaling In THP-1 Macrophages[J]. World Journal of Gastroenterology, 2016, 22(17): 4354-4361.
      样品类型: Cell culture supernatant
    28. 文献: Hao S Y, Ji J J, Zhao H T,et al. Mitochondrion-Targeted Peptide SS-31 Inhibited Oxidized Low-Density Lipoproteins-Induced Foam Cell Formation Through Both ROS Scavenging And Inhibition Of Cholesterol Influx In Raw264.7 Cells[J]. Molecules, 2015, 20(12): 21287-21297.
      样品类型: Cell lysate
    29. 文献: Veena V K, Popavath R N, Kennedy K, et al. In Vitro Antiproliferative, Pro-Apoptotic, Antimetastatic And Anti-Inflammatory Potential Of 2, 4-Diacteylphloroglucinol (DAPG) By Pseudomonas Aeruginosa Strain FP10[J]. Apoptosis, 2015, 20(10): 1281-1295.
      样品类型: Cell culture supernatant
    30. 文献: Tang Y, Zhang Y C, Chen Y, et al. The Role Of miR-19b In The Inhibition Of Endothelial Cell Apoptosis And Its Relationship With Coronary Artery Disease[J]. Scientific Reports, 2015.
      样品类型: Serum
    31. 文献: Cheng P, Hua W, Zhang W, et al. Decreased Monocyte Subsets And TLR4-Mediated Functions In Patients With Acute Severe Fever With Thrombocytopenia Syndrome (SFTS)[J]. International Journal of Infectious Diseases, 2015, 43: 37-42.
      样品类型: Cell culture supernatant
    32. 文献: Liao Y, Liu P, Guo F, et al. Oxidative Burst Of Circulating Neutrophils Following Traumatic Brain Injury In Human[J]. Plos One, 2013.
      样品类型: Plasma
    如有任何问题,请拨打电话 027-87879180 或填写 在线留言 我们会第一时间联系您。