Brand AVX TPSE226M035R0125 Low impedance tantalum capacitor, AVX 22

Electronic scale crystal oscillator, 3.2*2.5mm 3225, 16MHz (16.000MHZ), 12PF, 10PPM, 20PPM, 30PPM

Probe domestic double-head probe 038-BB-5.7L, the shape of the two ends of the needle is outside the pointed needle

Many customers have mentioned that although the ELISA experiment seems simple at first glance, it can be quite tricky in practice, leading to various issues that affect detection accuracy and test quality. Problems such as non-specific binding, false positives, background noise, and low signal-to-noise ratios often occur. These challenges require careful attention during the experimental process, continuous improvement, and a focus on key steps to ensure reliable results. First: Selection of Coating Solution The most commonly used coating buffer is carbonate buffer with a pH of 9.6. However, depending on specific testing requirements, other buffers like phosphate buffer or Tris-HCl (pH 7.2–8.0) may also be suitable. When using polystyrene plates, proteins bind through physical adsorption, which is non-specific. The strength of this adsorption depends on factors such as protein molecular weight, isoelectric point, and hydrophobic interactions. Smaller proteins tend to have more hydrophobic groups, making them easier to adsorb onto solid surfaces. This principle is crucial for optimizing the coating step and improving assay performance. Second: Blocking the Plate After coating, the next critical step is blocking, which involves filling unoccupied sites on the plate surface to prevent non-specific binding. Common blocking agents include 0.05%–0.5% bovine serum albumin (BSA), 10% or 1% gelatin, and skim milk powder. Skim milk is cost-effective and works well at higher concentrations (5%–10%). In some cases, animal sera are used to reduce cross-reactivity with similar proteins or casein. The choice of blocking agent should be based on the specific needs of the experiment and the type of sample being tested. Third: Washing the Plate Washing is one of the most important steps in an ELISA procedure. Since proteins tend to adsorb non-specifically onto plastic surfaces, thorough washing is essential to remove unbound enzymes, free ligands, and interfering substances. Incomplete washing can lead to high background signals and inaccurate results. Although automated washers are available, manual washing is still widely used, and human error can significantly impact outcomes. It’s important to follow standardized washing protocols and ensure all wells are properly rinsed. Fourth: Adding Antibodies (Primary and Secondary) When adding antibodies, it's important to handle the pipette carefully to avoid contamination or inaccurate dosing. The sample should be diluted appropriately, and the same dilution buffer should be used for both primary and secondary antibodies. If using two different antibodies, their working concentrations must be carefully optimized. Too high a concentration can cause excessive background, while too low a concentration may result in weak signals. Proper titration is essential for achieving optimal sensitivity and specificity.

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