Capture operation is a necessary condition to ensure accurate and reliable ELISA results - Database & Sql Blog Articles

High-quality reagents, proper instrumentation, and precise operation are all essential for achieving accurate and reliable ELISA results. The ELISA procedure can vary depending on the type of solid-phase carrier used. In domestic medical testing, plate-based methods are commonly employed. This article will focus on the key steps involved in the capture method. Following detailed instructions and strictly adhering to the recommended procedures ensures reliable outcomes. During testing with our ELISA kit, we found that specific IgM and IgG antibodies against certain antigens may coexist in serum, which can interfere with the detection of IgM antibodies. To address this issue, we adopted an IgM multi-capture method. This involves first immobilizing all IgM in the serum—both specific and non-specific—onto a solid phase, and then removing IgG before detecting the specific IgM. Here’s how we do it: (1) An anti-human IgM antibody is attached to a solid-phase carrier, forming a solid-phase anti-human IgM. After incubation, the plate is washed thoroughly. (2) Diluted serum samples are added. During the incubation, the IgM antibodies in the serum bind to the solid-phase anti-IgM. After washing, any impurities or other immunoglobulins are removed. (3) A specific antigen reagent is introduced. This binds only to the specific IgM present on the solid phase. The plate is washed again to remove unbound components. (4) An enzyme-labeled specific antibody is added, allowing it to react with the antigen bound to the solid phase. Another wash follows to remove excess reagents. (5) A substrate is added, causing a color change if specific IgM antibodies are present in the sample. A visible color indicates a positive result. Our ELISA kits use several common methods, including: I. Double Antibody Sandwich Assay for Antigen: - The first animal's antibody is adsorbed onto the solid phase. - Antigen is added, forming an antigen-antibody complex. - The second animal's antibody is added, creating a secondary complex. - An enzyme-labeled anti-antibody is introduced. - Substrate is added, and the amount of substrate degradation reflects the amount of antigen present. II. Direct Method for Antigen Detection: - Antigen is adsorbed onto the solid support. - An enzyme-labeled antibody is added, forming an antigen-antibody complex. - Substrate is added, and the degree of substrate breakdown corresponds to the antigen concentration. III. Indirect Method for Antibody Detection: - Antigen is coated onto the solid support. - Serum containing the target antibody is added, forming a complex. - An enzyme-labeled secondary antibody is introduced. - Substrate is added, and the resulting color change indicates the presence and quantity of the antibody. Advantages of our ELISA kits include: - High specificity: Extensive cross-reactivity studies were conducted to minimize errors from non-specific interactions. - High precision: Spike recovery and linear dilution tests ensure accuracy, with lower CV values than similar products. - Excellent repeatability: Consistent performance across batches ensures minimal variation between lots. - High reliability: Optimized reagents reduce the risk of false positives and false negatives. - Wide standard curve: Provides a broad detection range while maintaining data accuracy. - Cost-effective: Using imported RD reagents combined with domestic packaging offers high quality at a competitive price. Whether you're conducting routine diagnostics or research, our ELISA kits are designed to deliver consistent, accurate, and reliable results.

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