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What Are Human Proteases?
Human proteases are a diverse group of enzymes responsible for breaking down proteins by cleaving the peptide bonds between amino acids. This process, known as proteolysis, is fundamental to many physiological functions, including digestion, immune response, cell signaling, and the regulation of cell life and death. Proteases are classified based on their catalytic mechanisms into several major families: serine, cysteine, aspartic, metalloproteases, and threonine proteases. Each family exhibits unique substrate specificities and modes of action, enabling them to control distinct biological processes.
Proteases maintain cellular homeostasis and facilitate various biological processes. For example, serine proteases such as trypsin and chymotrypsin play a key role in the digestive process, breaking down proteins in food into absorbable peptides. In the immune system, proteases such as granzyme B help destroy infected or cancerous cells. In addition, proteases are involved in the regulation of blood clotting, wound healing and the activation of growth factors.
Dysregulation of human proteases has been linked to a variety of diseases. Overactive proteases can lead to tissue destruction, triggering diseases such as arthritis, cancer metastasis and chronic inflammation. Conversely, insufficient protease activity impairs processes such as digestion and immune defense, leading to disease.
Human Protease Panel at Creative Proteomics
Creative Proteomics offers the Human Protease 15 Plex Panel utilizing advanced Luminex xMAP technology (Multi-Analyte Profiling). We can simultaneously detect and quantify multiple protease inhibitors in a single sample. The technology utilizes color-coded microspheres coated with specific capture antibodies to bind target analytes. This approach enables high-throughput, multiplexed analyses with exceptional sensitivity and specificity.
The Luminex xMAP system is capable of handling complex samples and is compatible with a wide range of assay formats. This flexibility makes it an invaluable tool for researchers studying protease inhibitors in different biological contexts.
Detection Method
Magnetic bead-based Luminex multiplex assay
Species
Human
Analytes Detected
| Species | Specification | Protein Targets | Price |
|---|---|---|---|
| Human | Human Protease 15 Plex Panel | ADAMTS13, Cathepsin S, Kallikrein 3/PSA, Kallikrein 5, Kallikrein 6/Neurosin, MMP-1, MMP-2, MMP-3, MMP-8, MMP-9, MMP-12, MMP-13, Proprotein Convertase 9/PCSK9, Proteinase 3/Myeloblastin/PRTN3, u-Plasminogen Activator (uPA)/Urokinase | +Inquiry |
Advantages of the Human Protease Luminex Assay
- Multiplexing Capability: The ability to analyze multiple protease inhibitors simultaneously reduces sample volume requirements and increases throughput, making it highly efficient for large-scale studies.
- High Sensitivity and Specificity: The assay's design ensures precise detection of low-abundance inhibitors, which is crucial for identifying subtle changes in protease activity associated with disease.
- Cost-Effective: By enabling the analysis of multiple targets in a single run, the Luminex assay reduces costs associated with reagents, labor, and time.
- Broad Dynamic Range: The assay can detect a wide range of inhibitor concentrations, accommodating both low and high levels of protease activity in samples.
- Versatility: Compatible with various sample types, including serum, plasma, and cell culture supernatants, the Luminex assay is adaptable to different research needs.
Sample Requirements for Human Protease Luminex Panel
| Sample Type | Volume Required | Storage Conditions | Handling Instructions |
|---|---|---|---|
| Serum/Plasma | 50-100 µL | -80°C (long-term), -20°C (short-term) | Avoid repeated freeze-thaw cycles. Centrifuge to remove particulates. |
| Cell Lysates | 100-200 µg of protein | -80°C | Use protease inhibitors during lysis. Clarify lysates by centrifugation. |
| Tissue Homogenates | 100-200 µg of protein | -80°C | Homogenize in cold buffer with protease inhibitors. |
| Culture Supernatants | 500-1000 µL | -80°C | Filter to remove cells/debris. Store immediately after collection. |
What Can Human Proteases Do?
Understanding Disease Mechanisms
- Cancer: Protease inhibitors are crucial in controlling the activity of proteases involved in tumor progression and metastasis. Analyzing these inhibitors provides valuable insights into the underlying mechanisms of cancer development.
- cardiovascular diseases: Proteases contribute to the formation and rupture of atherosclerotic plaques, leading to heart attacks and strokes. By studying protease inhibitors, researchers can gain a deeper understanding of cardiovascular disease processes.
- Neurodegenerative Disorders: In neurodegenerative diseases such as Alzheimer's, protease inhibitors help regulate the breakdown of proteins, which can provide critical information on the factors driving neurodegeneration.
Advancing Drug Development
Protease inhibitors are essential in the drug discovery process, as they can be targeted to control specific proteases implicated in various diseases. Analyzing these inhibitors supports the development of new drugs and the optimization of existing therapies.
Biomarker Discovery
Protease inhibitors are important biomarkers for a wide range of diseases. Variations in their levels can indicate disease presence or progression. Through detailed analysis, new biomarkers can be discovered, enhancing diagnostic and monitoring capabilities.
Studying Immune Response
Protease inhibitors play a significant role in the regulation of immune responses. By analyzing these inhibitors, researchers can better understand how the immune system is modulated in both healthy and diseased states, providing insights into immune regulation and potential points of intervention.
Regulating Inflammatory Responses
Protease inhibitors help modulate inflammation by balancing the activity of proteases involved in inflammatory pathways. Understanding these inhibitors is critical for studying how inflammation is regulated in various conditions, aiding in the development of strategies to manage inflammation effectively.
In addition to preconfigured panels, we also offer customized analysis services. You can customize your own panel through our customization tool, or directly email us the targets you are interested in. A professional will contact you to discuss the feasibility of customization. We look forward to working with you!
What factors can influence the accuracy of the human protease inhibitor panel results?
Sample Quality: Poor sample quality, such as hemolysis or contamination, can affect the results. Proper collection, handling, and storage are critical.
Pre-analytical Variability: Differences in sample processing, such as varying centrifugation speeds or times, can introduce variability in the results.
Inhibitor Stability: Protease inhibitors may degrade over time or with repeated freeze-thaw cycles, leading to inaccurate measurements.
Assay Interference: The presence of interfering substances, such as lipids or other proteins, can impact assay performance, leading to skewed results.
How do you ensure the consistency and reproducibility of results in the human protease inhibitor panel?
Standardized Protocols: Adhering to strict protocols for sample preparation, handling, and processing minimizes variability.
Quality Controls: Including positive and negative controls in each assay run to monitor performance and ensure accuracy.
Calibration: Regular calibration of the Luminex xMAP technology and other equipment used in the assay to maintain precision.
Replicates: Running samples in replicates to verify consistency across measurements.
Can the human protease inhibitor panel be customized to include specific inhibitors of interest?
Yes, the human protease inhibitor panel can often be customized to include specific protease inhibitors of interest. Customization may involve adding additional targets to the panel or adjusting the assay to focus on particular protease inhibitors relevant to the customer's research or project needs. This flexibility allows for more targeted investigations and can be discussed with our scientific team to ensure the assay meets your specific requirements.
How can I determine if my research project would benefit from using the human protease inhibitor panel?
Determining the suitability of the Human Protease Panel for your research depends on several factors:
- Research Objectives: If your research involves studying protease regulation, proteolytic pathways, or the role of protease inhibitors in disease, this panel is highly relevant.
- Target Analytes: Review the specific protease inhibitors covered in the panel to ensure they align with your research focus.
- Sample Availability: Ensure that the types of samples you have are compatible with the panel's requirements and that you can meet the necessary volume and quality criteria.
- Data Needs: Consider if the multiplexing capabilities and detailed data provided by this panel would provide the insights you need for your project.
What steps are involved in the data analysis and interpretation of human protease inhibitor panel results?
The data analysis process typically involves:
- Data Acquisition: Raw data are collected from the Luminex xMAP system.
- Data Normalization: Normalizing the data to control for variability across samples and assay runs.
- Statistical Analysis: Applying statistical methods to determine the significance of the findings, often comparing against controls or baseline values.
- Interpretation: Analyzing the data in the context of the research question, considering the biological significance of any observed changes in protease inhibitor levels.
How should I prepare my samples if they have been stored for a long period before testing?
Thawing: Thaw samples slowly at 4°C to preserve the integrity of the protease inhibitors.
Aliquoting: If not already aliquoted, avoid repeated freeze-thaw cycles by dividing samples into smaller aliquots.
Inspection: Check for any signs of degradation, such as cloudiness or precipitation, before use.
Pre-analysis Processing: Follow any specific pre-analysis preparation steps, such as centrifugation or filtration, to remove potential contaminants or debris.
