The ProQuantum HumanVEGFImmunoassay Kit is designed to provide highly sensitive quantitative measurements of human VEGF 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VEGF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VEGF (vascular endothelial growth factor) which is a 45 kDa homodimeric, disulfide-linked glycoprotein involved in angiogenesis which promotes tumor progression and metastasis. VEGF has a variety of effects on vascular endothelium, including the ability to promote endothelial cell viability, mitogenesis, chemotaxis, and vascular permeability. The VEGF family currently includes VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, and PIGF. VEGF and its receptor system have been shown to be the fundamental regulators in the cell signaling of angiogenesis. Most tumors have the absolute requirement of angiogenesis, and VEGF has been described as the most potent angiogenic cytokine linked to this process. To date 5 different isoforms of VEGF have been described. These isoforms are generated as the result of alternative splicing from a single VEGF gene. These various isoforms have been shown to bind to two tyrosine-kinase receptors flt-1 (VEGFR-1) and flk-1/KDR (VEGFR-2), which have been found to be expressed almost exclusively on endothelial cells. VEGF and its high-affinity binding receptors, the tyrosine kinases FLK1 and FLT1, are thought to be important for the development of embryonic vasculature. Studies have shown that an alternately spliced form of FLT1 produces a soluble protein, termed sFLT1, which binds vascular endothelial growth factor with high affinity, playing an inhibitory role in angiogenesis. Elevated levels of VEGF is linked to POEMS syndrome (Polyneuropathy, Organomegaly, Endocrinopathy, Monoclonal gammopathy, Skin changes) also known as Crow-Fukase syndrome which affects multiple organs in the body.VEGF(血管内皮生长因子)是一种45 kDa的同型二聚体,二硫键连接的糖蛋白,参与血管生成,可促进肿瘤进展和转移。 VEGF对血管内皮具有多种作用,包括促进内皮细胞活力,促有丝分裂,趋化性和血管通透性的能力。 VEGF家族目前包括VEGF-A,VEGF-B,VEGF-C,VEGF-D,VEGF-E和PIGF。 VEGF及其受体系统已被证明是血管新生细胞信号传导中的基本调节剂。大多数肿瘤对血管生成具有绝对的要求,而VEGF被描述为与此过程相关的最有效的血管生成细胞因子。迄今为止,已经描述了5种不同的VEGF同工型。这些同工型是从单个VEGF基因进行选择性剪接的结果。已显示这些各种同工型与两个酪氨酸激酶受体flt-1(VEGFR-1)和flk-1 / KDR(VEGFR-2)结合,发现它们几乎只在内皮细胞上表达。 VEGF及其高亲和力结合受体酪氨酸激酶FLK1和FLT1被认为对胚胎脉管系统的发育很重要。研究表明,FLT1的另一种剪接形式可产生称为sFLT1的可溶性蛋白,该蛋白以高亲和力结合血管内皮生长因子,在血管生成中起抑制作用。 VEGF水平升高与POEMS综合征(多发性神经病,器官肿大,内分泌病,单克隆丙种球蛋白病,皮肤变化)有关,也被称为Crow-Fukase综合征,它会影响体内的多个器官。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。 |