High Sensitive ELISA Kit for Interleukin 35 (IL35)
- Product No.HEC008Hu
- 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 0.62pg/mL.
- Sample TypeSerum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- DownloadInstruction Manual
- UOM48T96T96T*596T*1096T*100
- FOBUS$ 539 For more details, please contact local distributors!US$ 770 For more details, please contact local distributors!US$ 3465 For more details, please contact local distributors!US$ 6545 For more details, please contact local distributors!US$ 53900 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 Interleukin 35 (IL35)
This assay has high sensitivity and excellent specificity for detection of High Sensitive Interleukin 35 (IL35).No significant cross-reactivity or interference between High Sensitive Interleukin 35 (IL35) and analogues was observed.
Recovery of the High Sensitive ELISA Kit for Interleukin 35 (IL35)
Matrices listed below were spiked with certain level of recombinant High Sensitive Interleukin 35 (IL35) and the recovery rates were calculated by comparing the measured value to the expected amount of High Sensitive Interleukin 35 (IL35) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 88-102 | 98 |
| EDTA plasma(n=5) | 98-105 | 101 |
| heparin plasma(n=5) | 78-88 | 81 |
Precision of the High Sensitive ELISA Kit for Interleukin 35 (IL35)
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Interleukin 35 (IL35) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Interleukin 35 (IL35) 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 Interleukin 35 (IL35)
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of High Sensitive Interleukin 35 (IL35) 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% | 87-101% | 78-105% | 85-102% |
| EDTA plasma(n=5) | 91-98% | 79-90% | 80-102% | 89-97% |
| heparin plasma(n=5) | 80-91% | 89-103% | 95-102% | 79-103% |
Stability of the High Sensitive ELISA Kit for Interleukin 35 (IL35)
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 Interleukin 35 (IL35)
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 Interleukin 35 (IL35)
The microplate provided in this kit has been pre-coated with a monoclonal antibody specific to IL12A. Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated monoclonal antibody specific to IL27B. 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 IL35, 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 IL35 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!) |
| RPC008Hu01 | Recombinant Interleukin 35 (IL35) | Positive Control; Immunogen; SDS-PAGE; WB. |
| APC008Hu01 | Active Interleukin 35 (IL35) | Cell culture; Activity Assays. |
| PAC008Hu01 | Polyclonal Antibody to Interleukin 35 (IL35) | WB; IHC; ICC; IP. |
| MAC008Hu21 | Monoclonal Antibody to Interleukin 35 (IL35) | WB; IHC; ICC; IP. |
| LAC008Hu72 | Biotin-Linked Monoclonal Antibody to Interleukin 35 (IL35) | WB; IHC; ICC. |
| SEC008Hu | ELISA Kit for Interleukin 35 (IL35) | Enzyme-linked immunosorbent assay for Antigen Detection. |
| HEC008Hu | High Sensitive ELISA Kit for Interleukin 35 (IL35) | Enzyme-linked immunosorbent assay for Antigen Detection. |
| PSC008Hu01 | Antibody Pair for Interleukin 35 (IL35) | ELISA; CLIA; ELISPOT; Luminex; Immunochromatography and other Immunoassays. |
| KSC008Hu01 | ELISA Kit DIY Materials for Interleukin 35 (IL35) | Main materials for "Do It (ELISA Kit) Yourself". |
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1、直接竞争，标记抗原，与检测样品中的抗原竞争抗体。
2、间接竞争，标记抗体，固相抗原与液相抗原（样品）竞争标记抗体。
3、定义
间接竞争法的模型：包被抗原，用HRP-抗体与样本一起加入。样本中的Ag与Solid-Ag竞争HRP-Ab，固相吸附的HRP-Ab与样本中的Ag浓度成反比。
直接竞争法的模型：包被抗体，用HRP-抗原与样本一起加入。样本中的Ag与HRP-Ag竞争Solid-Ab，固相吸附的HRP-Ag与样本中的Ag浓度成反比。
4、竞争法的理论基础：是限量抗体。只有在限量抗体基础上，两种抗原才会形成竞争关系。
5、、间接竞争法具备较高灵敏度原因。
直接竞争法里，标记抗原与待测抗原均是液相，与抗体的结合机会是一样的，例如有1份标记抗原与1份待测抗原竞争1份抗体，那么有50%的标记抗原能与抗体结合，所以标记抗原的相对结合率为50%。间接法里，固相抗原与抗体的接触面积较小，固相抗原与待测抗原的结合抗体机会是不平等的，接近顺序饱和法，即只有与待测抗原结合剩余的抗体才会与固相抗原结合，同样有1份固相抗原与1份待测抗原竞争1份抗体时，基本上抗体会被待测抗原中和掉，与固相抗原结合的抗体非常少。固相抗原的相对结合率为0%。因此，间接法的抑制曲线斜率会大于竞争法。
因为抑制率越大则斜率越大，从而灵敏度越大。（假设零管变异5%，以两倍SD为灵敏度限，则为90%相对结合率，则间接法可以在较低的待测抗原浓度达到这一相对结合率，因此灵敏度要高。）
在进行系统放大时，间接法一般可以使用酶标二抗。因为二抗可以针对抗体的多个部位，所以存在放大效应，从而能提高间接法的灵敏度。直接法一般难以进行放大，常用的有生物 素化抗原与酶标亲和素，但模式上似乎不存在放大效应。
6、间接法的高灵敏度难以实现的原因：
双抗体 夹心的免放（IRMA）模式刚出现时，也被模型证明灵敏度优于竞争法的放免（RIA），原因也是较大的斜率，但是IRMA的高灵敏度一直到单抗发展后才得以实现。
采用抗-HBe抗体包被反应板，加入校准品及被测样本，同时加入定量HBeAg中和抗原，经过振荡孵育,洗板后再加入铕标记的抗-HBe，若标本中抗-HBe浓度高，HBeAg将被大量中和，使最后形成的抗-HBe-HBeAg-铕标记抗-HBe复合物减少。增强液（β－NTA）将标记在抗体上的Eu3+解离到溶液中，Eu3+和增强液中的有效成分形成高荧光强度的螯合物，荧光强度和样本中的抗-HBe浓度成反比。
