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What is the relationship between the latest ELISA tips and the test reference?
Yesterday afternoon, in response to a customer's question, Mr. Wang Hengyuan, our technical commissioner, provided a detailed telephone explanation regarding the relationship between ELISA and test references. He emphasized that quantitative ELISA kits must include reference standards for generating standard curves, typically containing 4–5 concentrations that cover the detectable range. These standards are usually prepared in buffers with protein protectants and preservatives to ensure stability and accuracy.
During the discussion, Mr. Wang highlighted several key points:
First, the positive control. This is an essential component used to validate the experiment and serve as a benchmark for interpreting results. The composition of the reference material, especially the negative control, should closely resemble that of the actual sample being tested. For instance, when using human serum as a specimen, the control should ideally also be human serum. Normal human serum can produce varying levels of background depending on the ELISA format. At Shanghai Hengyuan Biotech Co., Ltd., we provide high-quality kits. However, because obtaining large quantities of normal human serum is challenging, many foreign ELISA kits use recalcified human plasma—where calcium ions are added to cause clotting, and the resulting liquid after removing the clot has a composition similar to serum.
Second, the negative control. This is used to confirm that the sample does not contain the target substance. For example, a negative control for HBsAg detection should not contain HBsAg and preferably also be negative for anti-HBs. The positive control is typically a buffer containing a protein stabilizer, with a known amount of the target substance added. This amount is usually specified in the kit’s instructions and should match the sensitivity of the reagent. By comparing the absorbance of the control with that of the sample, you can estimate the concentration of the target substance in the sample. For instance, the sensitivity of an ELISA kit for HBsAg detection is about 0.5 ng/ml, while the positive control usually contains around 10 ng/ml. Preservatives and antibiotics are often included in the controls to enhance shelf life.
**How to choose the right ELISA kit:**
1. Determine which proteins you need to detect.
2. Identify which species have the best homology with the gene encoding the target protein.
3. Look for kits designed for those species.
4. Check with the supplier to learn what type of antibodies are used in the kit.
5. Make your own decision based on the information gathered.
Choosing the right ELISA kit requires careful consideration of multiple factors, including the target protein, species specificity, and antibody quality. With the right tools and guidance, you can ensure accurate and reliable results every time.