当前位置：
How ProQuantum immunoasssays work
(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.
Recommended sample types are EDTA plasma and serum. A range of additional sample types are compatible with the technology (PEA), including citrate plasma, heparin plasma, cerebrospinal fluid, (CSF), tissue and cell lysates, fine needle biopsies, microdialysis fluid, cell culture media, dried blood spots, synovial fluid, saliva, plaque extract and microvesicles. Please note that protein expression levels are expected to vary in different sample types. Certain assays are differentially affected by interfering substances such as hemolysate. Download any of our Data Validation documents or contact support@olink.com for more information.
The calibrator curve shown below visualizes the analytical measuring range data based on in vitro measurement of recombinant antigen. Please note that this is shown for reference only and CANNOT be used to convert NPX values to absolute concentrations for proteins measured in plasma or serum samples. The vertical dotted lines represent LLOQ and ULOQ respectively, and the horizontal line indicates the LOD.
Components are color coded for ease of use.
产品属性/ Product Details
| 产品名称 | 人白介素6(IL-6)检测试剂盒 | 反应种属 | |
| 基因别名 | BSF-2, BSF2, CDF, HGF, HSF, IFN-beta-2, IFNB2, IL-6, IL6 | 基因ID | (Human) 3569 |
| Uniprot ID | P15692 | 蛋白别名 | B-cell differentiation factor, B-cell stimulatory factor 2, BSF-2, CDF, CTL differentiation factor, H-IL-6, ILN, Interleukin, Interleukin6, M-IL-6, R-IL-6, Hybridoma growth factor, IFN-beta-2, IL-6, Interferon beta-2, interleukin BSF-2, Interleukin-6 |
| 检测方法 | Proximity-based Immunoassay | 灵敏度 | <0.05 pg/mL |
| 检测仪器 | qPCR Instruments | 检测范围 | 0.064-10,000 pg/mL |
| 检测样本类型/体积 | Serum=2-5 µL, Supernatant=2-5 µL, Plasma=2-5 µL | ||
| 检测总时间 | 2 hr | 手工操作时间 | 30 min |
| 运输条件 | Dry ice | 储存温度 | -20°C |
| 均相(无需洗涤) | Yes | 标签或染料 | FAM™ |
| 关于本试剂盒 | The ProQuantum Human IL-6 Immunoassay Kit is designed to provide highly sensitive quantitative measurements of human IL-6 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. • High sensitivity-detect low levels of protein with greater sensitivity than traditional methods • Broad dynamic range-≥5 logarithmic units, minimizing sample dilutions to ensure they fall within the range • Small sample consumption-use 2-5 µL of sample (compared to 75 µL for triplicate wells with other methods) • Fast, easy workflow-2 hours from sample to answer, with no wash steps • No proprietary instrument to purchase-runs on any real-time PCR instrument 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 (Figure 1). 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. |
||
| 靶标信息 | IL-6 is a gene that encodes a cytokine that plays a role in inflammation and the maturation of B cells. The protein produced by this gene is an endogenous pyrogen that can induce fever in people with autoimmune diseases or infections. The gene is primarily expressed at sites of acute and chronic inflammation, where it is secreted into the serum and induces a transcriptional inflammatory response through interleukin 6 receptor, alpha. The functioning of this gene is implicated in a wide variety of inflammation-associated disease states, including suspectibility to diabetes mellitus and systemic juvenile rheumatoid arthritis. Elevated levels of the encoded protein have been found in virus infections, including COVID-19 (disease caused by SARS-CoV-2). Diseases associated with IL6 include Kaposi Sarcoma and Rheumatoid Arthritis, Systemic Juvenile. | ||
检测流程/ Assay Procedures
FAQs/ FAQs
Q1:请问血清样品稀释可以用DD水吗?
不能的，需要使用试剂盒自带的标准品&样品稀释液。
Q2:请问血清样品稀释可以用DD水吗?
不能的，需要使用试剂盒自带的标准品&样品稀释液。