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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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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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。
1.包被：用0.05MPH9.牰碳酸盐包被缓冲液将抗体稀释至蛋白质含量为1～10μg/ml。在每个聚苯乙烯板的反应孔中加0.1ml，4℃ 过夜。次日，弃去孔内溶液，用洗涤缓冲液洗3次，每次3分钟。（简称洗涤，下同）。
2. 加样：加一定稀释的待检样品0.1ml于上述已包被之反应孔中，置37℃ 孵育1小时。然后洗涤。（同时做空白孔，阴性对照孔及阳性对照孔）。
3. 加酶标抗体：于各反应孔中，加入新鲜稀释的酶标抗体（经滴定后的稀释度）0.1ml。37℃ 孵育0.5～1小时，洗涤。
4. 加底物液显色：于各反应孔中加入临时配制的TMB底物溶液0.1ml，37℃ 10～30分钟。
5. 终止反应：于各反应孔中加入2M硫酸0.05ml。
6. 结果判定：可于白色背景上，直接用肉眼观察结果：反应孔内颜色越深，阳性程度越强，阴性反应为无色或极浅，依据所呈颜色的深浅，以“+”、“-”号表示。也可测OD值：在ELISA检测仪上，于450nm（若以ABTS显色，则410nm）处，以空白对照孔调零后测各孔OD值，若大于规定的阴性对照OD值的2.1倍，即为阳性。
间接法
1.用包被缓冲液将已知抗原稀释至1～10μg/ml，每孔加0.1ml，4℃过夜；
2.次日洗涤3次；
3.加一定稀释的待检样品（未知抗体）0.1ml于上述已包被之反应孔中,置37℃孵育1小时,洗涤；
4.（同时做空白、阴性及阳性孔对照）于反应孔中，加入新鲜稀释的酶标第二抗体（抗抗体）0.1ml；
5.37℃孵育35-60分钟，洗涤；
6.’最后一遍用DDW洗涤。
其余步骤同“双抗体夹心法”的4、5、6。
第一次：包被抗体浓度为：8ug/ml,4,2,1,0.5,0.25,0为A,B,C,D,E,F,G
血清稀释度为1:100
酶标抗体稀释度为：1:1000,1:5000,1:10000,1:25000为1,2，3,4
结果为：1234
A3.3440.7120.4900.238
B2.7570.5230.3450.176
C2.4280.4550.3100.172
D>4.51.7771.2150.439
E>4.51.3570.7890.424
F1.1090.2540.1720.110
G1.3660.3310.1970.069(空白)从结果可以判断包被抗体浓度1ug/ml最合适
第二次：包被抗体1ug/ml
血清稀释度为1:25,1:50,1:100,1:150,1:200,1:250,1:300,空白为A,B,C,D,E,F,G,H
酶标抗体稀释度为：1:1000,1:5000,1:10000,1:25000为1,2，3,4
结果为：1234
A2.9930.6390.3380.236
B3.0210.7220.3070.191
C2.3360.4960.3080.176
D2.2560.4640.3040.176
E2.0310.4160.2470.139
F1.8300.3360.2310.153
G1.5980.3320.2270.132
空白H0.1580.0780.0720.074
这样的话应该是血清稀释度1:50最合适，酶标抗体稀释度1:1000最合适，但是这样的话OD值为3.021，是不是太大了？OD值是不是在1左右比较好呢？应该选择哪个浓度，哪个稀释度来做正式试验呢？
请大家帮帮忙，第一步加完抗原后必须封闭吗
1、重复性不好；
2、收自身抗体、嗜异性抗体等干扰，易出现假阳性；
3、不论仪器和手工操作，干扰因素较多。影响最大的是温度和时间。1、直接法（direct ELISA）将抗原直接固定在固相载体上，参加酶符号的一级抗体，即可测定抗原总量，此一级抗体的特异性非常重要。优势：操作手续简略，因无须运用二抗可防止交互反响。缺陷：实验中的一抗都得用酶符号，但不是每种抗体都适合做符号，费用相对进步。2.间接法（indirect ELISA）此测定办法与直接法相似，不一样在于一级抗体没有酶符号，改用酶符号的二级抗体去辨识一级抗体来测定抗原量。优势：二抗能够加强信号，并且有多种挑选能做不一样的测定剖析。不加酶符号的一级抗体则能保存它最多的免疫反响性。缺陷：交互反响发作的机率较高。3.双抗体夹心法（sandwich ELISA）被检测的抗原包被在两个抗体之间，其间一个抗体将抗原固定于固相载体上，即捕捉抗体。另一个则是检测抗体，此抗体可用酶符号后直接测定抗原的量；或不符号，再透过酶符号的二级抗体来测定抗原的量。这两种抗体有必要当心选择，才可防止交互反响或竞争一样的抗原联系部位。优势：高活络、高专一性，抗原无须事前纯化。缺陷：抗原必定得具有两个以上的抗体联系部位。4.竞争法（competitive ELISA）样本里的抗原（自在抗原）和纯化并固定在固相载体上的抗原（固定抗原）一同竞争一样的抗体，当样品里的自在抗原越多，就能够联系越多的抗体，而固定抗原就只能联系到较少的抗体，反之亦然。经清洁过程，洗去自在抗原和抗体的复合物，只留下固定抗原和抗体的复合物，拿来与只要固定抗原的对照组成果相比拟，依据呈色区别就可计算出样品里的抗原含量。优势：可适用比拟不纯的样本，并且数据再现性很高。缺陷：全体的敏感性和专一性都较差。本数据来源于百度地图，最终结果以百度地图最新数据为准。
