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| Product Name | High Density Lipoprotein Cholesterol (HDL-C) Content Assay Kit (Microplate Reader) |
| Catalog No. | BBITK-HMM-0022 |
| Description | High-density lipoprotein (HDL), as an anti-atherosclerotic lipoprotein, can transport cholesterol from extrahepatic tissues to the liver for metabolism, converting it into bile acids or directly excreting it from the intestine through bile, thereby reducing cholesterol deposition on the arterial walls. The anti-atherosclerotic effect is achieved through various means such as promoting reverse cholesterol transport, anti-inflammation, anti-oxidation, inhibiting thrombosis formation and improving endothelial cell function. The level of its content in the blood is of great significance for the risk assessment of cardiovascular diseases. |
| Testing Equipment | Microplate Reader |
| Matching | 96-well plate |
| Number of Testable Samples | 100 Samples |
| Estimated Measurement Time | 6 h (100 Samples) |
| Storage | Store at 4°C away from light |
| Self-contained Reagents | / |
| Detection Principle | The CHOD-PAP endpoint method combined with the classical GPO Trinder enzymatic reaction was used for the determination. Cholesterol esterase (CHE) was able to break down cholesterol esters into free cholesterol, and cholesterol oxidase (COD) further oxidized free cholesterol into cholestenone and H2O2, and H2O2 was able to react with 4-aminoantipyrine to produce red benzoquinone imine catalyzed by peroxidase (POD). H2O2 can be catalyzed by peroxidase (POD) to react with 4-aminoantipyrine to form red benzoquinone imine, and the product has a characteristic absorption peak at 550 nm, which can be used to quantitatively detect the HDL cholesterol content by the change of absorbance value. |
| Detection Methods | CHOD-PAP End Point Method |
| Detection Wavelength | 550 nm |
| Signal Response | Incremental |
| Standard | Cholesterol |
| Reference Standards | y=0.391x-0.0037 (R2=0.9992) |
| Standard Linear Range | 0.125-3.0 mmol/L |
| Detection Limit | 0.125 mmol/L |
| Note | If the A measurement or ΔA measurement exceeds the standard linear absorbance range: higher than the maximum value, it is recommended to appropriately dilute the sample to be tested with PBS or normal saline before conducting the measurement. If the sample size is lower than the minimum value, it is recommended to appropriately increase the sample size before conducting the measurement and make corresponding modifications during the calculation. This product adopts the HDL separation method and cholesterol determination method. It is simple and reliable to operate, has high sensitivity and good repeatability, and is not affected by the chemical substances in the vast majority of samples. However, in the system, vitamin C>0.18 g/L, hemoglobin >2 g/L, bilirubin >0.25 g/L, and strong reducing agents (such as dithiostachitol, mercaptoethanol, etc.) will interfere with the test results; Naturally coagulated serum and EDTA anticoagulated plasma can be used. Heparin is not recommended as an anticoagulant. |
High-density lipoprotein cholesterol (HDL-C) is a core research target in the fields of lipid metabolism, atherosclerosis, and cardiovascular disease (CVD) mechanisms. As a key component of lipoprotein research, HDL-C’s unique reverse cholesterol transport (RCT) function and anti-atherosclerotic properties have made it a focal point for life science researchers worldwide. With the deepening of studies on metabolic disorders, chronic diseases, and drug development, the demand for accurate, reliable, and research-oriented HDL-C detection tools has grown significantly in academic institutions, biotech laboratories, and pharmaceutical R&D teams.
Research Significance in Life Sciences
HDL-C is widely recognized as a critical marker in lipid metabolism research, as its ability to transport excess cholesterol from peripheral tissues to the liver for excretion directly impacts the development of atherosclerotic lesions. Studies have shown that dysregulated HDL-C levels or function are closely linked to the pathogenesis of metabolic syndrome, diabetes-related vascular complications, and CVDs—making HDL-C detection essential for verifying research hypotheses and exploring disease mechanisms.
In basic research, HDL-C serves as a key endpoint for evaluating the effects of genetic modifications, environmental factors, or experimental interventions. For example, researchers studying gene knockout/knock-in animal models (e.g., mice with altered apolipoprotein A1 expression) rely on precise HDL-C quantification to assess the impact of genetic changes on lipoprotein metabolism.
Limitations of Traditional Research Tools and Market Demand
Early HDL-C detection methods for research (such as ultracentrifugation and precipitation assays) are hindered by complex operation procedures, long processing times, and poor reproducibility. These methods often require large sample volumes and specialized equipment, making them unsuitable for high-throughput research projects or small-scale laboratory settings.
Modern enzymatic detection technologies, exemplified by the CHOD-PAP endpoint method adopted by this kit, have addressed these limitations. They offer high specificity, rapid reaction kinetics, and compatibility with standard laboratory equipment (microplate readers), meeting the core needs of researchers for efficient sample processing, consistent results, and scalability—whether for in vitro cell culture studies, animal model experiments, or large-scale epidemiological research samples.
Research Application Scenarios
Basic Lipid Metabolism Research: Used to quantify HDL-C levels in cell supernatants (e.g., hepatocytes, macrophages), animal serum/plasma (mice, rats, rabbits), or human research samples (de-identified specimens for epidemiological studies) to explore the regulatory mechanisms of HDL synthesis, secretion, and metabolism.
Atherosclerosis and CVD Mechanism Studies: Applied to analyze changes in HDL-C levels during plaque formation and progression, or to verify the role of HDL-C in inhibiting inflammation, oxidative stress, and thrombosis—key processes in atherosclerotic development.
Pharmaceutical and Biotech R&D: Utilized in preclinical drug development to evaluate the efficacy of potential lipid-modifying agents (e.g., novel HDL mimetics, cholesterol-lowering compounds). Researchers can monitor HDL-C levels in animal models or in vitro systems to assess drug-induced changes and optimize dosage regimens.
Nutrition and Environmental Research: Employed to study the impact of dietary interventions (e.g., omega-3 fatty acid supplementation, plant sterol intake) or environmental factors (e.g., pollution, lifestyle modifications) on HDL-C metabolism in experimental models.
Research-Grade Specificity: Combines the classical GPO Trinder enzymatic reaction with the CHOD-PAP endpoint method. Cholesterol esterase (CHE) specifically hydrolyzes cholesterol esters into free cholesterol, and cholesterol oxidase (COD) targets free cholesterol for oxidation—ensuring that only HDL-bound cholesterol is detected without cross-reactivity with other lipoproteins (LDL, VLDL) or common research sample components.
Optimized Linear Detection Range: Covers 0.125-3.0 mmol/L, which aligns with the HDL-C concentration range in most research samples (e.g., animal serum, cell culture supernatants, and human research specimens). This range eliminates the need for frequent sample dilution or concentration, streamlining experimental workflows.
Sensitive Detection Limit: Features a minimum detectable concentration of 0.125 mmol/L, enabling accurate quantification of low HDL-C levels in samples such as those from gene-edited animal models with impaired HDL synthesis or in vitro experiments with limited sample volumes.
Laboratory-Friendly Operation: Compatible with standard 96-well plates and common microplate readers, supporting simultaneous detection of up to 100 samples per kit. The protocol is straightforward with clear step-by-step instructions, requiring no specialized technical training—ideal for busy research laboratories with diverse experimental needs.
Stable Storage Conditions: Can be stored at 4°C away from light, avoiding the need for ultra-low temperature storage (-20°C/-80°C) that is required for some research kits. This simplifies inventory management and reduces reagent degradation risks during transportation or long-term storage.
High Research Data Reliability: The reference standard curve (y=0.391x-0.0037) has a correlation coefficient (R²) of 0.9992, indicating excellent linearity and quantitative accuracy. This ensures that experimental results are consistent and reproducible—critical for publishing research findings and validating experimental conclusions.
Time-Efficient for Research Workflows: The estimated measurement time for 100 samples is only 6 hours, which is 30%-50% faster than traditional methods (e.g., ultracentrifugation, precipitation assays). This efficiency allows researchers to process more samples in less time, accelerating project progress.
Flexible Sample Compatibility: Supports testing of naturally coagulated serum and EDTA-anticoagulated plasma—two of the most commonly used sample types in life science research. This flexibility eliminates constraints on sample collection and processing, adapting to diverse experimental designs.
High-Throughput Compatible: The 96-well plate format integrates seamlessly with automated liquid handling systems and high-throughput microplate readers, making it suitable for large-scale research projects (e.g., drug screening libraries, epidemiological research cohorts) that require processing hundreds of samples.
For research use only, not for clinical use.
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