ELISA Kit for Transforming Growth Factor Beta 1 (TGFb1)
TGF-B1; CED; DPD1; LAP; Camurati-Engelmann Disease; Latency-associated peptide
- Product No.CEA124Bo
- Organism SpeciesBos taurus; Bovine (Cattle) Same name, Different species.
- All
- Human
- Mouse
- Rat
- Cavia
- Rabbit
- Simian
- Caprine
- Ovine
- Equine
- Bovine
- Porcine
- Gallus
- Canine
- Others
- Multi-species
- Pan-species
- Test MethodCompetitive Inhibition
- Assay Length2h
- Detection Range12.35-1000pg/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 4.67pg/mL.
- Sample Typeserum, platelet-poor plasma, tissue homogenates, cell culture supernates and other biological fluids
- DownloadInstruction Manual
- UOM48T96T96T*596T*1096T*100
- FOBUS$ 494 For more details, please contact local distributors!US$ 706 For more details, please contact local distributors!US$ 3177 For more details, please contact local distributors!US$ 6001 For more details, please contact local distributors!US$ 49420 For more details, please contact local distributors!
Packages (Simulation)- Packages (Simulation)
- Results demonstration
- Typical Standard Curve
- ISO9001: 2008, ISO13485: 2003 Registered
Specificity of the ELISA Kit for Transforming Growth Factor Beta 1 (TGFb1)
This assay has high sensitivity and excellent specificity for detection of Transforming Growth Factor Beta 1 (TGFb1).No significant cross-reactivity or interference between Transforming Growth Factor Beta 1 (TGFb1) and analogues was observed.
Recovery of the ELISA Kit for Transforming Growth Factor Beta 1 (TGFb1)
Matrices listed below were spiked with certain level of recombinant Transforming Growth Factor Beta 1 (TGFb1) and the recovery rates were calculated by comparing the measured value to the expected amount of Transforming Growth Factor Beta 1 (TGFb1) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 92-101 | 97 |
| EDTA plasma(n=5) | 85-101 | 97 |
| heparin plasma(n=5) | 91-104 | 97 |
Precision of the ELISA Kit for Transforming Growth Factor Beta 1 (TGFb1)
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Transforming Growth Factor Beta 1 (TGFb1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Transforming Growth Factor Beta 1 (TGFb1) 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 ELISA Kit for Transforming Growth Factor Beta 1 (TGFb1)
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Transforming Growth Factor Beta 1 (TGFb1) 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) | 89-104% | 89-97% | 90-97% | 86-101% |
| EDTA plasma(n=5) | 86-104% | 90-97% | 93-105% | 78-98% |
| heparin plasma(n=5) | 89-97% | 99-105% | 95-103% | 85-104% |
Stability of the ELISA Kit for Transforming Growth Factor Beta 1 (TGFb1)
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 ELISA Kit for Transforming Growth Factor Beta 1 (TGFb1)
1. Prepare all reagents, samples and standards;2. Add 50µL standard or sample to each well.And then add 50µL prepared Detection Reagent A immediately.Shake and mix. Incubate 1 hour at 37°C;3. Aspirate and wash 3 times;4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;5. Aspirate and wash 5 times;6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;7. Add 50µL Stop Solution. Read at 450 nm immediately.
Test principle of the ELISA Kit for Transforming Growth Factor Beta 1 (TGFb1)
This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Transforming Growth Factor Beta 1 (TGFb1) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Transforming Growth Factor Beta 1 (TGFb1) and unlabeled Transforming Growth Factor Beta 1 (TGFb1) (Standards or samples) with the pre-coated antibody specific to Transforming Growth Factor Beta 1 (TGFb1). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Transforming Growth Factor Beta 1 (TGFb1) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Transforming Growth Factor Beta 1 (TGFb1) in the sample.
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Single-component Reagents of Assay KitLysis Buffer Specific for ELISA / CLIAQuality Control of ELISA / CLIAELISA Kit Customized ServiceDisease Model Customized ServiceSerums Customized ServiceTGFB1 Activation ReagentReal Time PCR Experimental Service
Related products
| Catalog No. | Organism species: Bos taurus; Bovine (Cattle) | Applications (RESEARCH USE ONLY!) |
| RPA124Bo01 | Recombinant Transforming Growth Factor Beta 1 (TGFb1) | Positive Control; Immunogen; SDS-PAGE; WB. |
| PAA124Bo01 | Polyclonal Antibody to Transforming Growth Factor Beta 1 (TGFb1) | WB; IHC; ICC; IP. |
| MAA124Bo21 | Monoclonal Antibody to Transforming Growth Factor Beta 1 (TGFb1) | WB; IHC; ICC; IP. |
| SEA124Bo | ELISA Kit for Transforming Growth Factor Beta 1 (TGFb1) | Enzyme-linked immunosorbent assay for Antigen Detection. |
| CEA124Bo | ELISA Kit for Transforming Growth Factor Beta 1 (TGFb1) | 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。
