Research on zebrafish transforming growth factor beta (TGF - β) ELISA detection kit

Shanghai Enzyme Bio Technology Co., Ltd. (brand name: MEIAOBIO) is a high-tech enterprise dedicated to the field of life sciences, committed to providing high-quality and innovative biological reagents and solutions for global scientific research institutions, medical institutions, and industrial users. Relying on the advantages of Shanghai as an international center for scientific and technological innovation, the company has gathered top-notch technical teams and R&D resources in the industry, and with a professional, accurate, and reliable service concept, helps with life science research and industrial development. MEIAOBIO's core product areas cover four major directions: molecular biology, cell biology, immunoassay, and biological sample library. The main products include ELISA kits, which are highly sensitive and specific detection tools widely used in the analysis of disease biomarkers, cytokines, cytokines, and other factors. Biochemical reagent kit: covering metabolic pathways, enzyme activity detection and other fields, supporting basic research and industrial quality inspection needs. PCR Kit: Provides a fast and accurate nucleic acid amplification solution, suitable for gene cloning, pathogen detection, and expression analysis. Biological sample library products: including cell lines, standardized serum, microbial strains, and biochemical reagents, ensuring experimental reproducibility and data reliability. The company's technical strength and quality assurance are driven by technological innovation and has established a strict quality control system. At the same time, MEIAOBIO maintains close cooperation with domestic and foreign research institutions and universities, continuously optimizes product performance, promotes technological iteration, and ensures the provision of cutting-edge and stable experimental tools for users. Service Philosophy MEIAOBIO adheres to the service tenet of "customer-centric", providing customized experimental solutions, professional technical guidance, and efficient logistics support. From product selection to after-sales tracking, we provide customers with one-stop solutions throughout the process, helping to improve research efficiency and transform achievements. Vision and Mission: In the future, MEIAOBIO will continue to deeply cultivate the field of life sciences, empower global scientific research and medical undertakings with innovative technologies, promote the progress of precision medicine, biomedicine and other industries, and contribute to human health and social development. Brand positioning: A professional, trustworthy, and innovative provider of biotechnology solutions. Brand commitment: Quality first, service based, to assist every scientific exploration!
Research on zebrafish transforming growth factor beta (TGF - β) ELISA detection kit

1、 Core components of ELISA kit

ELISA kits typically contain the following key components, and different types of kits (such as sandwich methods, indirect methods, etc.) may have slight differences:

1. Coated Plate: A 96 well (mostly made of polystyrene) surface coated with specific antibodies (or antigens) for capturing target molecules in the sample.
2. Standards/calibrators: Target molecules of known concentration (such as recombinant antigens, purified antibodies) used to draw standard curves and quantify the concentration of target molecules in the sample.
3. Detection of antibodies: Specific antibodies against target molecules (often labeled with enzymes such as HRP horseradish peroxidase and AP alkaline phosphatase).
4. Enzyme conjugate diluent: a buffer used to dilute enzyme labeled antibodies.
5. Substrate solution: Enzyme catalyzed substrates (such as TMB colorimetric solution and OPD colorimetric solution corresponding to HRP); The PNPP colorimetric solution corresponding to AP reacts to produce colored products.
6. Termination solution: Terminate the enzymatic reaction and fix the color development result.
7. Washing solution (PBST or TBST): used to wash away unbound non-specific substances and reduce background interference.
8. Sealing liquid: After coating, seal the unbound sites of the microporous plate to prevent non-specific adsorption.
9. Sample diluent: used to dilute the test sample (such as serum, plasma, tissue homogenate, etc.) and match the reaction system.

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2、 The detection principle of ELISA

The core of ELISA is the coupling of antigen antibody specific binding and enzymatic colorimetric reaction. The main steps are as follows (taking the "sandwich method" as an example):

1. Coating: Coating specific antibodies (capture antibodies) onto the surface of a microplate to form solid-phase antibodies.
2. Sealing: Add sealing solution and fill the unbound sites of the antibody to avoid non-specific adsorption of impurities in the sample.
3. Sample addition: Add the test sample (including the target antigen) or standard substance, and the antigen in the sample binds to the solid-phase antibody.
4. Incubation and washing: Incubate at room temperature or 37 ℃ to fully bind the antigen antibody; Wash to remove unbound substances.
5. Add detection antibody: Add enzyme labeled detection antibody (which binds to another epitope of the target antigen) to form a "solid-phase antibody antigen enzyme labeled antibody" complex (sandwich structure).
6. Wash again: Remove unbound enzyme-linked antibodies.
7. Color development with substrate addition: Adding substrate solution, enzymes (such as HRP) catalyze the substrate to generate colored products (color intensity is positively correlated with the concentration of the target antigen).
8. Termination reaction: Add termination solution (such as sulfuric acid) to stop the color reaction.
9. Reading analysis: Use an enzyme-linked immunosorbent assay (ELISA) reader to measure the absorbance (OD) at a specific wavelength (such as 450 nm for HRP), and calculate the concentration of the target antigen in the sample using a standard curve.

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3、 The main types of ELISA

According to the different detection targets (antigens/antibodies) and reaction principles, ELISA can be divided into the following categories:

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4、 Operation precautions

To ensure the accuracy and reproducibility of the test results, the following key points should be noted:

1. Sample processing: Avoid hemolysis, lipid blood or lysozyme interference; The sample needs to be diluted according to the instructions.
2. Incubation conditions: Strictly control the temperature (usually 37 ℃) and time (30 minutes to 2 hours) to avoid temperature fluctuations or insufficient/excessive time.
3. Washing operation: Insufficient washing can lead to high background, and excessive washing may wash away the bound substances; Suggest using a washing machine (set 5 washes, each for 30 seconds).
4. Color control: The substrate should be stored away from light, and the color development time should be adjusted according to the sample concentration.
5. Control settings: Blank control, negative control, positive control, and standard samples must be set up to verify the effectiveness of the experiment.

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5、 Quality Control and Interpretation of Results

1. Standard curve: Draw a standard curve (usually an S-shaped curve) with the concentration of the standard sample as the horizontal axis and the OD value as the vertical axis, and calculate the concentration corresponding to the OD value of the sample (pay attention to the linear range, and dilute or concentrate the sample if it exceeds the range).
2. Sensitivity and specificity: Sensitivity (lowest detection limit) and specificity (cross reactivity with analogues) are the core indicators for evaluating the performance of a reagent kit, which should be referred to in the instructions or verified through pre experiments.
3. Data validity: If the R ² of the standard curve is less than 0.99, the negative control OD value is too high (>0.1), or the positive control does not meet the standard, the experiment needs to be repeated.

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6、 Application Fields

• Research field: For scientific research only.

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7、 Suggestions for choosing ELISA kits

• Clear detection target: is it an antigen (such as viral protein) or an antibody (such as IgG/IgM)? Small molecules (such as drugs) require the use of competitive assay kits.
• Sample type: serum, plasma, tissue homogenate or cell supernatant? Select a matching sample diluent.
• Performance requirements: Quantitative testing should focus on linear range and repeatability; Qualitative testing can focus on setting the cut-off value.

Research on zebrafish transforming growth factor beta (TGF - β) ELISA detection kit


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