Mini Samples ELISA Kit for Interleukin 3 (IL3)
MCGF; MULTI-CSF; MCSF; M-CSF; Multiple; Multilineage-Colony-Stimulating Factor; Hematopoietic Growth Factor; P-Cell Stimulating Factor; Mast-Cell Growth Factor
- Product No.MEA076Mu
- Organism SpeciesMus musculus (Mouse) 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 Rangen/a
- Sensitivityn/a
- Sample Typen/a
- Downloadn/a
- UOM48T96T96T*596T*1096T*100
- FOBUS$ 545 For more details, please contact local distributors!US$ 778 For more details, please contact local distributors!US$ 3501 For more details, please contact local distributors!US$ 6613 For more details, please contact local distributors!US$ 54460 For more details, please contact local distributors!
Packages (Simulation)- Packages (Simulation)
- Results demonstration
- ISO9001: 2008, ISO13485: 2003 Registered
Specificity of the Mini Samples ELISA Kit for Interleukin 3 (IL3)
This assay has high sensitivity and excellent specificity for detection of Mini Samples Interleukin 3 (IL3).No significant cross-reactivity or interference between Mini Samples Interleukin 3 (IL3) and analogues was observed.
Recovery of the Mini Samples ELISA Kit for Interleukin 3 (IL3)
Matrices listed below were spiked with certain level of recombinant Mini Samples Interleukin 3 (IL3) and the recovery rates were calculated by comparing the measured value to the expected amount of Mini Samples Interleukin 3 (IL3) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 92-101 | 97 |
| EDTA plasma(n=5) | 85-99 | 93 |
| heparin plasma(n=5) | 96-104 | 101 |
Precision of the Mini Samples ELISA Kit for Interleukin 3 (IL3)
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mini Samples Interleukin 3 (IL3) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mini Samples Interleukin 3 (IL3) 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 Mini Samples ELISA Kit for Interleukin 3 (IL3)
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Mini Samples Interleukin 3 (IL3) 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) | 90-103% | 82-103% | 84-91% | 78-97% |
| EDTA plasma(n=5) | 88-95% | 90-97% | 81-101% | 79-97% |
| heparin plasma(n=5) | 97-104% | 80-102% | 97-105% | 87-94% |
Stability of the Mini Samples ELISA Kit for Interleukin 3 (IL3)
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 Mini Samples ELISA Kit for Interleukin 3 (IL3)
1. Prepare all reagents, samples and standards;2. Add 25µL standard or sample to each well. Incubate 1 hour at 37°C;3. Aspirate and add 25µL prepared Detection Reagent A. Incubate 1 hour at 37°C;4. Aspirate and wash 3 times;5. Add 25µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;6. Aspirate and wash 5 times;7. Add 25µL Substrate Solution. Incubate 10-20 minutes at 37°C;8. Add 20µL Stop Solution. Read at 450nm immediately.
Test principle of the Mini Samples ELISA Kit for Interleukin 3 (IL3)
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 Mini Samples Interleukin 3 (IL3). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Mini Samples Interleukin 3 (IL3). 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 Mini Samples Interleukin 3 (IL3), 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 Mini Samples Interleukin 3 (IL3) 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: Mus musculus (Mouse) | Applications (RESEARCH USE ONLY!) |
| RPA076Mu02 | Recombinant Interleukin 3 (IL3) | Positive Control; Immunogen; SDS-PAGE; WB. |
| RPA076Mu01 | Recombinant Interleukin 3 (IL3) | Positive Control; Immunogen; SDS-PAGE; WB. |
| PAA076Mu02 | Polyclonal Antibody to Interleukin 3 (IL3) | WB; IHC; ICC; IP. |
| PAA076Mu01 | Polyclonal Antibody to Interleukin 3 (IL3) | WB; IHC; ICC; IP. |
| MEA076Mu | Mini Samples ELISA Kit for Interleukin 3 (IL3) | Enzyme-linked immunosorbent assay for Antigen Detection. |
| SEA076Mu | ELISA Kit for Interleukin 3 (IL3) | Enzyme-linked immunosorbent assay for Antigen Detection. |
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2.加大辣根过氧化物酶标记的链霉亲和素；
我现在用商品化的试剂盒进行夹心法ELISA测定某抗体的浓度，盒子给定了4个标准品，标准品抗体浓度为70000~1000单位，测定步骤中要求对样品进行1:1000稀释后测定。
但是有的样本按照1:1000稀释后，最终OD值大于70000浓度的标准品的OD值，然后使用ELISACALC软件进行四参数拟合，超过了标准曲线的范围，就算不出来这个样品的浓度，但是如果在1000-70000之间的OD值就可以算出相应浓度。请教大神们，接下来我是否可以对样品进行1:2000,1:4000浓度稀释，算出结果后再×2、×4，算作此样品的浓度呢？还是直接给一个＞70000的定性结果？或者可以有能算出来的其他软件？
Ps试剂盒中提及Dilutionlinearity（稀释线性？）为141%，这个是什么意思呢？
多谢！！
请大家帮帮忙，第一步加完抗原后必须封闭吗
目前，我用该蛋白的鼠源性单抗来包被ELISA板子，5%BSA37度封闭二小时后，洗涤四次后将板子分三个组加入抗原：其中一个组中加入的是我重组的蛋白，另一个组为用其它方法证实含该蛋白的人体内血清，最后一个组加入PBS。37度一小时后洗涤四次后每孔加入我制备的兔源性多抗。37度一小时洗涤六次后我再加入酶标记的抗兔的抗体37度40分钟后洗涤六次后TMB显色。
问题就在我显色后所有的孔都是阳性，其中以抗原为我的重组蛋白组与PBS组OD值最高，两组都可以达到1.8以上，而加入的抗原为人体血清组低，为0.6-1.0不等。
后面我优化条件，封闭液用过5%的脱脂奶粉，2%的BSA，包被的单抗浓度摸了梯度，加入的多抗也摸了梯度，酶标的抗体也摸了梯度。结果都没有改变，连趋势每次都是一样以重组蛋白组与PBS组OD最高。
今天我还试着把抗原换成其它蛋白也做出了强阳性？
请教各位高手，我的ELISA该如何往下面做了啊，是不是我的抗体本身有问题呢？
对了，我包被用的单抗是用GE公司的预装柱纯化的，很纯，在考染中是看不到一条杂带的。
ELISA双抗体夹心法(enzyme linked immunosorbent assay——sandwich technique)的原理是将特异性抗体结合到固相载体上形成固相抗体，然后和待检血清中的相应抗原结合形成免疫复合物，洗涤后再加酶标记抗体，与免疫复合物中抗原结合形成酶标抗体-抗原-固相抗体复合物，加底物显色，判断抗原含量。
生物帮有相关介绍。编码RNA http://doc.bio1000.com/show-3399.html
（2）封闭：将已包被的板用洗涤液洗涤2次，每孔加入120 μL封闭液，封闭一段时间，取出，甩干备用。
（3）竞争反应：在制备好的板中每孔加入50 μL系列浓度的标准品溶液和50 μL稀释好的酶标抗体溶液，孵育一段时间。
（4）洗板：用洗涤液洗涤5次。
（5）显色反应：每孔加入显色溶液100 μL，孵育一段时间。
（6）终止：每孔各加入终止液，在酶标仪上测定各孔的吸光值。
