Key Biological Reagents by ANT BIO PTE. LTD.: Hyaluronidase & Enzyme-Free Sterile Water for Biomedical Research
In the fields of biomedicine and cell research, high-quality biological reagents are the cornerstone of successful experiments. Among them, hyaluronidase and enzyme-free sterile water are two indispensable core reagents with distinct functions yet equally critical roles. Hyaluronidase, as a "molecular scissors" for extracellular matrix processing, and enzyme-free sterile water, as the "source of life" for molecular biology experiments, provide strong support for various research work. This article systematically introduces the definitions, mechanisms of action, multi-field applications, and experimental schemes of these two reagents.
1. Definition and Mechanism of Action: The Key to Precise Cleavage
Hyaluronidase is a general term for a family of enzymes capable of degrading hyaluronic acid. Hyaluronic acid is a linear macromolecular polysaccharide widely present in the extracellular matrix of animal tissues, connective tissues, and the capsules of certain bacteria. As one of the main components of the extracellular matrix, it has high hydrophilicity and viscoelasticity, which can form a hydrated gel to support, retain water and protect cells. Meanwhile, it also serves as a physical barrier for cell migration, proliferation, and signal transduction.
The mechanism of action of hyaluronidase lies in its ability to specifically hydrolyze the β-1,4-glycosidic bonds linking N-acetyl-D-glucosamine and D-glucuronic acid in hyaluronic acid molecules. Through this cleavage, hyaluronidase can decompose the huge hyaluronic acid long chains into oligosaccharide fragments, thereby achieving the following effects:
• Reducing Tissue Viscosity: Destroying the gel-like barrier formed by hyaluronic acid.
• Increasing Tissue Permeability: Creating channels for the diffusion and migration of other molecules, drugs, or cells.
• Dissociating Cells from the Matrix: Helping to release cells embedded in the matrix.
It is based on this core mechanism that hyaluronidase excels in multiple research and application fields.
2. Overview of Main Applications
The applications of hyaluronidase mainly revolve around its ability to "deconstruct" hyaluronic acid, which can be divided into the following major directions:
• Tissue Dissociation: Used to digest tissues rich in hyaluronic acid when preparing primary cells, softening them and releasing cells.
• Assisted Drug Delivery: As a diffusing agent, it promotes the absorption and distribution of subcutaneously or intramuscularly injected drugs and reduces local swelling.
• Cell Culture and Analysis: Used to treat certain cells that secrete a large amount of extracellular matrix during cell passage or harvesting.
• Reproductive Biology Research: Used to treat the cell layer around oocytes in assisted reproductive technology.
• Basic Research: Used to study the specific functions of hyaluronic acid in cell behavior, development, and diseases (such as cancer and inflammation).
3. Experimental Application Scenarios and Operational Considerations
The following elaborates on the application scenarios, key points of scheme design, and precautions of hyaluronidase in specific experiments.
Application Scenario: Isolating viable and functional primary cells from animal or human tissues (such as skin, umbilical cord, cartilage, tumor tissue, etc.).
Experimental Protocol:
• Tissue Pretreatment: Cut the tissue into small pieces (1-3 mm³).
• Enzymatic Digestion: Soak the tissue pieces in a digestion solution containing hyaluronidase (usually with a working concentration of 0.5 - 2 mg/mL). To improve digestion efficiency, hyaluronidase is often used in combination with other hydrolases such as collagenase and trypsin to form a composite enzyme digestion solution, which synergistically decomposes different components in the extracellular matrix.
• Incubation and Termination: Incubate with continuous shaking at 37°C for 30 minutes to several hours, depending on the type and thickness of the tissue. Regularly sample and observe the cell dissociation under a microscope. Once fully dissociated, immediately add medium containing serum (serum contains enzyme inhibitors) to terminate the reaction.
• Cell Collection: Filter through a cell sieve to remove undigested tissue blocks, centrifuge to collect cells, resuspend for counting and culture.
Precautions:
• Concentration and Time Optimization: Excessively high enzyme concentration or too long digestion time will damage the cell membrane, reducing cell viability and adhesion ability. Pre-experiments must be performed to determine the optimal conditions.
• Aseptic Operation: The enzyme solution needs to be filtered and sterilized, and the entire operation process should be carried out in a ultra-clean workbench.
• Activity Verification: The activity of enzymes from different batches or sources may vary, so attention should be paid to their specific activity units.
Application Scenario: Certain cell lines (such as some cancer cells or mesenchymal stem cells) secrete abundant extracellular matrix during culture, forming a dense cell layer. When using conventional trypsin digestion, the cells fall off in sheets and are not completely dissociated.
Experimental Protocol:
• Before trypsin digestion, the cells can be pretreated with a buffer containing hyaluronidase (for example, 0.1 - 0.5 mg/mL) for 5-15 minutes.
• Hyaluronidase first degrades hyaluronic acid in the matrix, weakening the connection between cells and the matrix, and then adding trypsin can more gently and efficiently dissociate the cells into a single cell suspension, which is conducive to subsequent counting, passage, or seeding.
3.3 Assisted Reproductive Technology Research
Application Scenario: In in vitro fertilization experiments, it is necessary to remove the cumulus cells and corona radiata around the oocytes to observe and evaluate oocyte maturity or perform micromanipulation (such as ICSI).
Experimental Protocol:
• Place the collected cumulus-oocyte complexes in a culture medium containing hyaluronidase (usually a lower concentration is used, such as 80-100 IU/mL).
• Act at 37°C for 30 seconds to several minutes, and observe closely. Once the cumulus cells spread out, immediately wash the oocytes with culture medium multiple times to completely remove the enzyme solution and shed granulosa cells.
• This method can quickly and relatively gently expose the oocytes, avoiding physical damage that may be caused by mechanical methods.
3.4 Disease Model and Mechanism Research
Application Scenario: Studying the role of hyaluronic acid in tumor invasion, metastasis, angiogenesis, and inflammatory response.
Experimental Protocol:
• Loss-of-Function Research: Add hyaluronidase locally in the cell culture system or animal models (such as chick chorioallantoic membrane model) to degrade local hyaluronic acid.
• Observation Indicators: By comparing the treatment group with the control group, the following can be observed:
￮ Cell invasion/migration ability: Use Transwell or scratch test to see if the invasion ability of cancer cells is weakened after degrading hyaluronic acid.
￮ Angiogenesis: Observe the promotion or inhibition of neovascularization after degrading hyaluronic acid.
￮ Drug permeability: Study whether degrading hyaluronic acid in the tumor matrix can enhance the penetration efficiency of chemotherapeutic drugs.
Such research provides direct evidence for clarifying the function of hyaluronic acid in physiological and pathological processes.
Hyaluronidase and enzyme-free sterile water, as key biological reagents in biomedical research, play irreplaceable roles in different experimental links. Hyaluronidase, as an efficient and specific biocatalyst, helps researchers break through the extracellular matrix barrier, laying a foundation for primary cell preparation and disease mechanism exploration. Enzyme-free sterile water provides a pure and safe experimental environment for sensitive molecular biology experiments, ensuring the reliability of nucleic acid-related research results. Mastering the proper use of these two reagents will open up more possibilities for scientific research. ANT BIO PTE. LTD. is committed to providing high-quality versions of these reagents to support researchers worldwide.
5. Recommended Hyaluronidase Products by ANT BIO PTE. LTD.
ANT BIO PTE. LTD. provides high-quality hyaluronidase products that have undergone strict quality control and verification, with stable activity and reliable performance, which can fully meet the needs of various scientific research experiments. The detailed product information is shown in the following table:
|
Cat. No. |
Product Name |
Specification |
|
abs42225221 |
Hyaluronidase |
10mg/50mg |
|
abs47014926 |
Hyaluronidase (Ovine Testes) |
100mg/1g |
|
abs44063396 |
Hyaluronidase (Ovine Testes) |
100mg/1g |
|
abs44063397 |
Hyaluronidase (Bovine Testes) |
500mg |
|
abs5510894 |
Human Hyaluronidase (HAase) ELISA Kit |
96T |
|
abs5520181 |
Mouse Hyaluronidase (HAase) ELISA Kit |
96T |
ANT BIO PTE. LTD. is committed to advancing life science research by providing high-quality, reliable reagents and comprehensive solutions. Our hyaluronidase products, as an important part of the molecular interaction research series, are carefully developed and optimized to meet the diverse needs of researchers in extracellular matrix-related experiments and emerging research fields.
Guided by the principles of innovation, quality, and customer-centricity, our three specialized sub-brands (Absin, Starter, and UA) cover a full spectrum of research needs, from general reagents to specialized experimental kits. We strive to establish long-term and trusted partnerships with researchers worldwide, supporting them in achieving breakthroughs in life science research and contributing to the development of fields such as cell biology, reproductive biology, and oncology.
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ANT BIO PTE. LTD. – Empowering Scientific Breakthroughs
At ANTBIO, we are committed to advancing life science research through high-quality, reliable reagents and comprehensive solutions. Our hyaluronidase products, with stable activity and strict quality control, provide strong support for various research fields such as primary cell isolation, cell culture, and disease mechanism exploration. Backed by professional technical support, we strive to be your trusted partner in unlocking scientific mysteries. Explore our product portfolio today and elevate your research to new heights.
For more product details, please visit: AntBio - Fueling Research, Feeding Discovery
9. Enzyme-Free Sterile Water by ANT BIO PTE. LTD.: The "Source of Life" for Molecular Biology Experiments
In molecular biology laboratories, there is a seemingly ordinary yet crucial reagent — enzyme-free sterile water. Although it appears simple, it is far more complex than ordinary distilled water. Its purity directly determines the success or failure of key experiments such as RNA extraction and cDNA synthesis. This specially treated water ensures that sensitive biomolecules are not interfered with by degrading enzymes, making it an indispensable basic reagent in molecular biology experiments.
9.1 What is Enzyme-Free Sterile Water?
Simply put, enzyme-free sterile water is high-purity water that has undergone special treatment to remove all nucleases and proteases while ensuring a sterile state. It is not ordinary distilled water or deionized water but has gone through more rigorous processing procedures and quality control.
According to different processing methods, enzyme-free sterile water is mainly divided into two categories: DEPC-treated water and non-DEPC-treated water.
• DEPC-Treated Water: Prepared by treating ultrapure deionized water with diethyl pyrocarbonate (DEPC) followed by autoclaving. This method inactivates enzymes through chemical modification, especially effectively removing RNase.
• Non-DEPC-Treated Water: Prepared by physical methods such as multiple distillation, autoclaving, and filtration sterilization, without containing DEPC components.
Both types of water ensure the absence of nuclease (DNase and RNase) and protease activity, making them suitable for the most sensitive molecular biology experiments.
9.2 Core Value of Enzyme-Free Sterile Water
In molecular biology experiments, trace amounts of contaminating enzymes can lead to the failure of the entire experiment. Especially RNase, which is extremely stable and ubiquitous, with a large amount present on the skin of experimental personnel.
The core value of enzyme-free sterile water lies in providing a pure "safe space" for sensitive experiments. Compared with laboratory-made pure water, commercial enzyme-free sterile water offers consistent quality and reliable performance, saving researchers the time and cost of processing water and verifying quality themselves.
For high-level research requiring publication or reproducible experimental results, this consistency in quality is indispensable.
9.3 Application Scenarios of Enzyme-Free Sterile Water
RNA molecules are particularly susceptible to degradation by RNase, so enzyme-free sterile water is especially important in RNA-related experiments. During the dissolution of RNA precipitates, the use of enzyme-free sterile water can prevent RNA from being degraded at the moment of dissolution.
Experiments such as reverse transcription reactions, in vitro transcription, and siRNA annealing all require water without RNase activity. In these experiments, the purity of water is directly related to the integrity of RNA molecules and the reliability of experimental results.
9.3.2 DNA Analysis Technologies
Although DNA is relatively more stable than RNA, the presence of DNase can also lead to DNA degradation and affect experimental results. In experiments such as PCR and cDNA synthesis, enzyme-free sterile water is used to prepare reaction systems, which can prevent the degradation of nucleic acid templates and products.
Technologies that require high nucleic acid integrity, such as molecular cloning, DNA labeling, and sequencing, also rely on enzyme-free sterile water to maintain the stability of DNA molecules.
9.3.3 DEPC-Sensitive Applications
In certain special cases, such as oocyte microinjection, residual DEPC may be toxic to cells. At this time, non-DEPC-treated enzyme-free sterile water should be selected.
Some sophisticated enzymology studies also tend to use non-DEPC-treated water to avoid the subtle effects that DEPC may have on the activity of certain enzymes.
9.3.4 Preparation of High-Standard Solutions
When preparing various high-purity solutions such as cell culture solutions, electrophoresis buffers, and enzyme storage buffers, the use of enzyme-free sterile water can avoid the introduction of impurities or enzyme contamination. Especially enzyme-free sterile water with low endotoxin content is more suitable for cell culture and related bioprocesses.
• For most RNA experiments, DEPC-treated water is the first choice because it can effectively inactivate RNase.
• For DEPC-sensitive applications, non-DEPC-treated enzyme-free sterile water should be selected.
• For routine DNA experiments and general molecular biology applications, non-DEPC-treated nuclease-free water can be selected, which is usually more economical.
• For cell experiments, especially sensitive cell lines, consideration should be given to using pyrogen-free and endotoxin-free enzyme-free sterile water.
• When using enzyme-free sterile water, disposable gloves must be worn, because there is usually RNase on the hands, and bare-hand operation may lead to contamination.
• If the amount used each time is small, it can be properly aliquoted before use to avoid contamination caused by repeated uncapping.
• Use sterile operation technology; even after filtration sterilization, attention should be paid to preventing microbial contamination.
• Pay attention to the product storage conditions and validity period; generally, it can be stored at room temperature, and the validity period is usually 12-24 months.
9.5 Recommended Enzyme-Free Sterile Water Products by ANT BIO PTE. LTD.
ANT BIO PTE. LTD. provides high-quality enzyme-free sterile water products that have undergone strict quality control and verification, with stable performance and reliable quality, which can fully meet the needs of various molecular biology experiments. The detailed product information is shown in the following table:
|
Cat. No. |
Product Name |
Specification |
|
abs9259 |
Enzyme-Free Sterile Water |
500mL |
