TNF alpha Human ProQuantum Immunoassay Kit/人TNF alpha ProQuantum免疫检测试剂盒
货号：A35601,A356015
规格：96 Tests,5X96 Tests
价格：6561,34927
产品类型：Proquantum ELISA
品牌：Thermo Fisher
物种：人
宿主：其它
抗体亚型：其它
荧光染料：其它
灵敏度 | <0.01 pg/mL |
| 检测范围 | 0.0128 – 5000 pg/mL |
样本类型/体积 | 血浆 2-5 μL 血清 2-5 μL 上清液 2-5 μL |
操作时间 | 30 min |
测试时间 | 2 hr |
Homogenous (no wash) | Yes |
仪器设备 | qPCR Instruments |
规格 | 5 X 96 Tests |
组份 | - 分析稀释缓冲液 - 抗体结合物 A - 抗体结合物 B - 抗体结合稀释缓冲液 - 标准品 - 预混料 - 连接酶 |
储存温度 | -20℃ |
| 运输条件 | 干冰 |
| Protein name | TNF alpha |
Protein aliases | tumor necrosis factor |
物种（Tested） | 人 |
试剂盒类型 | Proximity-based Immunoassay |
标记（染料） | FAM |
| Gene ID | (Human) 7124 |
| Gene aliases | DIF, TNF-alpha, TNFA, TNFSF2, TNLG1F |
| Gene symbol | TNF |
UniProt ID | (Human) P01375 |
| The ProQuantum HumanTNF alphaImmunoassay Kit is designed to provide highly sensitive quantitative measurements of human TNF alpha protein in small sample volumes. Utilizing proximity ligation assay (PLA) technology, the assay combines the analyte specificity of high-affinity antibody-antigen binding with the signal detection and amplification capabilities of real-time PCR to achieve a simple yet powerful next-generation protein quantitation platform. A user-friendly workflow combined with intuitive Cloud-based software for analytics enables sample-to-answer in just 2 hours.ProQuantum 免疫检测试剂盒进行少量人TNF alpha蛋白的高灵敏度定量检测。 利用邻近连接测定（PLA）技术，该测定将高亲和力抗体-抗原结合的分析物特异性与实时PCR的信号检测和扩增功能结合在一起，从而实现了一个简单而强大的下一代蛋白质定量平台。 友好的工作流程与直观的基于云的分析软件相结合，可在2小时内完成从样品到答案的分析。ProQuantum immunoassays utilize a matched pair of target-specific antibodies, each conjugated to a DNA oligonucleotide. During antibody-analyte binding, the two DNA oligos are brought into close proximity, which allows for ligation of the two strands and subsequent creation of a template strand for amplification. This platform leverages the sensitivity and large dynamic range of Applied Biosystems TaqMan real-time PCR technology.ProQuantum免疫测定法使用一对配对的靶标特异性抗体，每种抗体均与DNA寡核苷酸偶联。 在抗体-分析物结合过程中，两个DNA寡核苷酸紧密靠近，这允许两条链连接并随后创建模板链进行扩增。 该平台利用了Applied Biosystems TaqMan实时PCR技术的灵敏度和宽动态范围。The assay workflow is fast and easy—2 steps in 2 hours. There are a total of 7 components in each kit (Figure 2). First, mix the antibody-conjugates, dilute the sample, and create the standard curve in a working plate. Then, using a multi-channel pipette, add the antibody-conjugates and sample (or standard) into the wells of a PCR plate and incubate for 1 hour. Combine the master mix and ligase and add to the wells of the PCR plate, then run the plate on any qPCR instrument. After the run is complete, import the results file into the ProQuantum cloud-based software at https://apps.thermofisher.com/apps/proquantum. Using this software, the data can be analyzed easily to obtain protein concentration values. The software allows you to set up standard curves, design plate layouts, set up customized assay instructions, and obtain robust statistical group-wise comparisons.测定工作流程快速简便— 2小时内完成2个步骤。 每个试剂盒中总共有7个组件。 首先，混合抗体-结合物物，稀释样品，并在工作板上创建标准曲线。 然后，使用多通道移液器，将抗体结合物和样品（或标准品）添加到PCR板的孔中，并孵育1小时。 合并预混液和连接酶，并添加到PCR板的孔中，然后在任何qPCR仪器上运行该板。 运行完成后，将结果文件导入https://apps.thermofisher.com/apps/proquantum的基于ProQuantum云的软件中。 使用此软件，可以轻松分析数据以获得蛋白质浓度值。 该软件允许您设置标准曲线，设计板布局，设置定制化验说明并获得可靠的逐组统计比较。Target information 靶标信息Tumor Necrosis Factor is a polypeptide cytokine produced by monocytes and macrophages. It functions as a multipotent modulator of immune response and further acts as a potent pyrogen. TNF-alpha circulates throughout the body responding to stimuli (infectious agents or tissue injury), activating neutrophils, altering the properties of vascular endothelial cells, regulating metabolic activities of other tissues, as well as exhibiting tumoricidal activity by inducing localized blood clotting. TNF-alpha also inhibits lipoprotein lipase activity resulting in cachexia, a physical wasting condition. Activation of B-cells by the Epstein Barr virus can be inhibited by TNF-alpha. Due to its varied actions throughout the immune system, TNF-alpha may play a role in the pathogenesis of many disease states. TNF-alpha production is mediated by the action of lymphokines and endotoxins on the macrophage. Purified monocytes produce TNF-alpha within four hours of stimulation by recombinant IL-2 and there is some in vitro evidence to suggest that TNF-alpha is expressed at high levels and with prolonged kinetics in T cells stimulated by both CD2 and CD28. Secretion of TNF-alpha is enhanced by gamma interferon. TNF then induces or enhances the specific production of Class I MHC antigen, GM-CSF, and IL-1.肿瘤坏死因子是单核细胞和巨噬细胞产生的多肽细胞因子。它起着免疫反应多能调节剂的作用，并进一步起着有效的热原作用。 TNF-α在全身循环，对刺激（传染原或组织损伤）作出反应，激活中性粒细胞，改变血管内皮细胞的特性，调节其他组织的代谢活性，并通过诱导局部凝血而表现出杀肿瘤活性。 TNF-α还抑制脂蛋白脂肪酶的活性，导致恶病质，一种物理上的浪费状态。爱泼斯坦巴尔病毒对B细胞的激活可以被TNF-α抑制。由于其在整个免疫系统中的不同作用，TNF-α可能在许多疾病状态的发病机理中起作用。 TNF-α的产生是通过巨噬细胞上淋巴因子和内毒素的作用介导的。纯化的单核细胞在重组IL-2刺激的四个小时内产生TNF-α，并且有一些体外证据表明TNF-α在CD2和CD28刺激的T细胞中高水平表达，并且动力学延长。 γ-干扰素可增强TNF-α的分泌。然后，TNF诱导或增强I类MHC抗原，GM-CSF和IL-1的特异性产生。For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.仅供研究使用。 不用于诊断过程。 未经明确授权不得转售。 |
| 特点： |
| - 高灵敏度 - 更高的灵敏度检测低水平的蛋白质- 宽动态范围- 样品消耗少 - 使用2–5 µL样品- 快速，轻松的工作流程 - 从样品到取样仅需2个小时，无需清洗- 无需购买专有仪器 - 可在任何实时PCR仪器上运行 |
| 数据： |
| TNF alpha Human ProQuantum Immunoassay Kit<b>How ProQuantum immunoasssays work</b><br />(A) Antibody-antigen binding: Antibodies bind to two separate epitopes on the antigen (during a 1-hr incubation), which brings the two conjugated oligonucleotides into close proximity. (B) Ligation and amplification of signal (in a qPCR instrument): DNA ligase and a third splint oligonucleotide connector are added to ligate the two ends of the conjugated oligonucleotides, creating a 100-base DNA template. Once the ligase is inactivated at 95°C, the sample is amplified through 40 cycles of annealing and extension. The amount of DNA produced is measured after each amplification cycle via fluorescent dyes, which exhibit fluorescence that is directly proportional to the number of PCR product molecules (or amplicons) generated. This graph shows the fluorescence vs. cycle number curves for four different starting protein concentrations. The dashed line represents the fluorescence threshold. The cycle number required to reach the fluorescence threshold (threshold cycle or Ct) is plotted vs. protein concentration to create a standard curve.<b> ProQuantum免疫测定的工作原理</ b> <br />（A）抗体-抗原结合：抗体与抗原上的两个独立表位结合（在1小时的孵育过程中），这使两个结合的寡核苷酸紧密相邻。 （B）信号的连接和扩增（在qPCR仪器中）：添加DNA连接酶和第三个夹板寡核苷酸连接物以连接缀合的寡核苷酸的两端，从而产生100个碱基的DNA模板。连接酶在95°C失活后，样品会通过40个退火和延伸循环进行扩增。在每个扩增循环后，通过荧光染料测量产生的DNA量，该荧光染料显示的荧光与产生的PCR产物分子（或扩增子）的数量成正比。该图显示了四种不同起始蛋白浓度的荧光与循环数曲线。虚线表示荧光阈值。将达到荧光阈值所需的循环数（阈值循环或Ct）对蛋白质浓度作图，以创建标准曲线。 TNF alpha Human ProQuantum Immunoassay Kit<b>ProQuantum kit contents</b><br />Components are color coded for ease of use. |
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