High Sensitive ELISA Kit for Tumor Necrosis Factor Alpha (TNFa)
DIF; TNF-A; TNFSF2; Cachectin; Tumor Necrosis Factor Ligand Superfamily Member 2
- Product No.HEA133Hu
- Organism SpeciesHomo sapiens (Human) Same name, Different species.
- All
- Human
- Mouse
- Rat
- Cavia
- Rabbit
- Simian
- Caprine
- Ovine
- Equine
- Bovine
- Porcine
- Gallus
- Canine
- Others
- Multi-species
- Pan-species
- Test MethodDouble-antibody Sandwich
- Assay Length3h
- Detection Range1.56-100pg/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 0.52pg/mL.
- Sample Typeserum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- DownloadInstruction Manual
- UOM48T96T96T*596T*1096T*100
- FOBUS$ 413 For more details, please contact local distributors!US$ 590 For more details, please contact local distributors!US$ 2655 For more details, please contact local distributors!US$ 5015 For more details, please contact local distributors!US$ 41300 For more details, please contact local distributors!
Packages (Simulation)- Packages (Simulation)
- Results demonstration
- Typical Standard Curve
- ISO9001: 2008, ISO13485: 2003 Registered
Specificity of the High Sensitive ELISA Kit for Tumor Necrosis Factor Alpha (TNFa)
This assay has high sensitivity and excellent specificity for detection of High Sensitive Tumor Necrosis Factor Alpha (TNFa).No significant cross-reactivity or interference between High Sensitive Tumor Necrosis Factor Alpha (TNFa) and analogues was observed.
Recovery of the High Sensitive ELISA Kit for Tumor Necrosis Factor Alpha (TNFa)
Matrices listed below were spiked with certain level of recombinant High Sensitive Tumor Necrosis Factor Alpha (TNFa) and the recovery rates were calculated by comparing the measured value to the expected amount of High Sensitive Tumor Necrosis Factor Alpha (TNFa) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 88-96 | 91 |
| EDTA plasma(n=5) | 82-95 | 88 |
| heparin plasma(n=5) | 80-101 | 86 |
Precision of the High Sensitive ELISA Kit for Tumor Necrosis Factor Alpha (TNFa)
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Tumor Necrosis Factor Alpha (TNFa) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Tumor Necrosis Factor Alpha (TNFa) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10%>Inter-Assay: CV<12%>
Linearity of the High Sensitive ELISA Kit for Tumor Necrosis Factor Alpha (TNFa)
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of High Sensitive Tumor Necrosis Factor Alpha (TNFa) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Sample | 1:2 | 1:4 | 1:8 | 1:16 |
| serum(n=5) | 98-105% | 79-99% | 81-102% | 97-105% |
| EDTA plasma(n=5) | 83-104% | 85-101% | 82-92% | 80-98% |
| heparin plasma(n=5) | 95-103% | 83-95% | 85-96% | 79-103% |
Stability of the High Sensitive ELISA Kit for Tumor Necrosis Factor Alpha (TNFa)
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Assay procedure summary of the High Sensitive ELISA Kit for Tumor Necrosis Factor Alpha (TNFa)
1. Prepare all reagents, samples and standards;2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;4. Aspirate and wash 3 times;5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;6. Aspirate and wash 5 times;7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;8. Add 50µL Stop Solution. Read at 450nm immediately.
Test principle of the High Sensitive ELISA Kit for Tumor Necrosis Factor Alpha (TNFa)
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to High Sensitive Tumor Necrosis Factor Alpha (TNFa). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to High Sensitive Tumor Necrosis Factor Alpha (TNFa). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain High Sensitive Tumor Necrosis Factor Alpha (TNFa), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of High Sensitive Tumor Necrosis Factor Alpha (TNFa) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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Related products
| Catalog No. | Organism species: Homo sapiens (Human) | Applications (RESEARCH USE ONLY!) |
| APA133Hu01 | Active Tumor Necrosis Factor Alpha (TNFa) | Cell culture; Activity Assays. |
| RPA133Hu01 | Recombinant Tumor Necrosis Factor Alpha (TNFa) | Positive Control; Immunogen; SDS-PAGE; WB. |
| PAA133Hu01 | Polyclonal Antibody to Tumor Necrosis Factor Alpha (TNFa) | WB; IHC; ICC; IP. |
| LAA133Hu81 | FITC-Linked Polyclonal Antibody to Tumor Necrosis Factor Alpha (TNFa) | WB; IHC; ICC; IF. |
| LAA133Hu71 | Biotin-Linked Polyclonal Antibody to Tumor Necrosis Factor Alpha (TNFa) | WB; IHC; ICC. |
| MAA133Hu22 | Monoclonal Antibody to Tumor Necrosis Factor Alpha (TNFa) | WB; IHC; ICC; IP. |
| MAA133Hu21 | Monoclonal Antibody to Tumor Necrosis Factor Alpha (TNFa) | WB; IHC; ICC; IP. |
| LAA133Hu72 | Biotin-Linked Monoclonal Antibody to Tumor Necrosis Factor Alpha (TNFa) | WB; IHC; ICC. |
| MEA133Hu | Mini Samples ELISA Kit for Tumor Necrosis Factor Alpha (TNFa) | Enzyme-linked immunosorbent assay for Antigen Detection. |
| SEA133Hu | ELISA Kit for Tumor Necrosis Factor Alpha (TNFa) | Enzyme-linked immunosorbent assay for Antigen Detection. |
| HEA133Hu | High Sensitive ELISA Kit for Tumor Necrosis Factor Alpha (TNFa) | Enzyme-linked immunosorbent assay for Antigen Detection. |
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| SCA133Hu | CLIA Kit for Tumor Necrosis Factor Alpha (TNFa) | Chemiluminescent immunoassay for Antigen Detection. |
| PSA133Hu01 | Antibody Pair for Tumor Necrosis Factor Alpha (TNFa) | ELISA; CLIA; ELISPOT; Luminex; Immunochromatography and other Immunoassays. |
| KSA133Hu01 | ELISA Kit DIY Materials for Tumor Necrosis Factor Alpha (TNFa) | Main materials for "Do It(ELISA Kit) Yourself" |
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需要的仪器有：酶标仪、恒温箱、枪、枪头、烧杯、滤纸.当然有自动洗板机最好.
现在国内最差也是用3代试剂，有些地方会用4代试剂。
4代试剂（检查抗原+抗体）——窗口期为4周。因为抗原于3-4周达到复制的峰值，此时通过4代试剂检查，如果感染了HIV，抗原/抗体至少有一个为阳性，如果都是阴就排除了。
3代试剂（只查抗体）——窗口期为6周。
以上为理论分析+临床经验的结果，可以说是99.9%的准确度。
但是目前FDA、CDC和试剂生产商统一达成的共识，也就是针对普通人，最保守的窗口期是3个月。无论什么试剂，3个月都100%排除。
1．取出试剂盒室温平衡30min，取出血样放至室温。
2．配标准品：取150uL标准品加入150uL标准品稀释液稀释，依次稀释5次。
3．加样：分别于各反应孔中加入标准品50uL，样品40UL，标准品做复孔，样品做3孔。
4．分别于样品孔中加入10UL抗体。
5．标准品和样品孔中分别加入50uL链酶亲和素-HRP，盖上封板膜,轻轻震荡混匀，37℃温育60min。
6．配洗涤液：将30倍浓缩洗涤液用蒸馏水30倍稀释后备用。
7．洗涤：小心揭开封板膜，弃去液体，甩干，每孔加200uL洗涤液，静置30s后弃去，如此重复5次，拍干。
8．显色：每孔先加入显色剂A 50uL，再加显色剂B 50uL，轻轻震荡混匀，37℃避光显色6min。
9． 终止：每孔加入终止液50uL，终止反应（此时蓝色立即转为黄色）。
10．测定：以空白孔调零，450nm波长依序测量各孔的吸光度。测定应在加终止液10min之内进行。
11．保存结果，收拾桌面。
12．分析处理数据
注意事项
1. 取出板条前恢复到室温后再打开外包装袋，实验中不用的板条立即放回包装中，密闭封口，其余不用试剂应盖好。
2. 实验操作中请使用一次性的吸头，避免交叉污染。
3．实验板孔加入试剂的顺序应一致，以保证所有反应孔的孵育时间一致。
4．洗涤过程中反应孔中残留的洗涤液应在滤纸上充分拍干，勿将滤纸直接放入反应孔中吸水。
5．试剂盒内试剂请在保质期内使用，不同批号试剂不要混用。
6．1000pg/ml以上的结果为非线性的，根据此标准曲线无法得到精确的结果。大于1000pg/ml 的样品应以标准稀释缓冲液稀释后重做。在结果分析时，结合考虑相应的稀释度。
复习了很多文献在测定方法里面均提到了CV
例如：
Theintra-assaycoefficientofvariationfortheassaywas4-6%.
Theanalyticcoefficientofvariationwas10.2%
问题是：
我是否可以直接使用说明书里的CV值？
如果不可以，我该如何操作进行计算
