0.25% Trypsin-EDTA Solution
0.25% Trypsin-EDTA Solution is a sterile-filtered liquid cell dissociation reagent containing trypsin and EDTA for detaching adherent cells in culture. Supplied as a 500 mL frozen liquid, CCM-RG-0043 is formulated with 2.5 g/L trypsin and 0.38 g/L EDTA·4Na for routine passaging, scale-up support, and R&D workflows. This animal-origin reagent ships on dry ice, with free worldwide shipping available for qualified orders through CellCultureMedia.
- ✓ Free worldwide shipping
- ✓ ISO-aligned manufacturing
- ✓ Certificate of Analysis on request
- ✓ Bulk & custom volumes quoted on request
Overview
CCM-RG-0043 is a 0.25% Trypsin-EDTA Solution designed for controlled dissociation of adherent mammalian cell monolayers from culture vessels, microcarriers, and related growth surfaces. The formulation combines 2.5 g/L trypsin, a proteolytic enzyme used to cleave adhesion-associated proteins, with 0.38 g/L EDTA·4Na, a chelating agent that helps reduce calcium- and magnesium-dependent cell attachment. This dual-action composition supports consistent detachment when users need a standard animal-origin digestion reagent for cell culture operations.
The product is supplied as a sterile-filtered liquid in a 500 mL format for laboratory and process-development use. Key release specifications include pH 7.2–8.0 and osmolality 270–320 mOsmol/kg, supporting compatibility with common mammalian cell culture workflows when used according to an established protocol. The reagent is categorized under cell digestion and dissociation reagents within our cell culture reagents portfolio, making it suitable for procurement teams standardizing media-adjacent consumables across R&D, pilot, and production-support laboratories.
Applications
- Routine passaging of adherent cell cultures where enzymatic detachment is required before reseeding, counting, cryopreservation, or assay setup.
- Process-development studies evaluating harvest timing, exposure duration, neutralization strategy, and post-dissociation cell recovery across defined vessel formats.
- Scale-up workflows involving flasks, multilayer culture vessels, roller bottles, or microcarrier-based culture systems, subject to user qualification for the target cell type.
- R&D protocols requiring EDTA-assisted dissociation to improve detachment efficiency compared with enzyme-only approaches in calcium- and magnesium-sensitive attachment systems.
- Preparation of single-cell suspensions for downstream in vitro assays, transfection workflows, cell banking activities, or analytical sampling where a standardized detachment reagent is specified.
Because trypsin sensitivity varies by cell line, culture age, confluency, matrix coating, and serum-free or serum-containing conditions, users should validate exposure time and neutralization method before routine implementation. For high-value or shear-sensitive cultures, a small-scale qualification run is recommended to define detachment performance, morphology, viability, and reseeding behavior under site-specific conditions. Procurement managers can reference CCM-RG-0043 when requesting repeat lots, while technical teams can align incoming inspection with pH, osmolality, sterility, storage, and shelf-life requirements.
Storage and Handling
Store CCM-RG-0043 frozen at -30 to -5 °C and protected from light. The stated shelf life is 24 months when the product remains unopened and stored under recommended conditions. The product ships on dry ice to maintain the frozen state in transit; upon receipt, inspect the container, verify labeling, and transfer promptly to qualified frozen storage. Avoid repeated freeze-thaw cycles because enzymatic performance can decline after unnecessary thermal stress. For routine use, thaw only the required volume under controlled conditions, mix gently after thawing, and use aseptic technique to reduce contamination risk. Do not warm for extended periods, and return unused aliquots to appropriate storage only if your internal procedure permits refreezing. As with other enzymatic dissociation reagents, users should confirm that temperature, contact time, and quenching conditions are compatible with the specific culture system. For multi-site supply planning or larger lot requirements, submit details through request a quote so our team can coordinate availability, shipment timing, and documentation needs.
Quality and Documentation
CCM-RG-0043 is produced as a sterile-filtered, animal-origin liquid reagent for cell culture-related applications. Release information includes pH range, osmolality range, form, sterility method, storage condition, shipping condition, and shelf life, supporting technical review by quality, procurement, and process-development teams. A Certificate of Analysis is available on request for lot-specific documentation, and customers may use our quality information to support supplier qualification, incoming material review, and internal change-control files. Manufacturing and documentation practices are aligned with ISO-style quality expectations for research and bioprocessing supply chains, with attention to traceability, specification control, and consistent packaging.
CellCultureMedia supports international B2B ordering for academic, biotech, and biopharma laboratories. This 500 mL unit is priced at $201.39, and free worldwide shipping is available where order conditions and destination logistics permit. Dry-ice shipment is used for temperature control, and lead time may depend on inventory status, export routing, and cold-chain service availability. For recurring demand, users can request bulk pricing, scheduled supply, or reserved inventory to reduce sourcing variability across passaging, cell banking, and development campaigns.
Specifications
Formulation Highlights
Contains (+)
2.5 g/L trypsin, 0.38 g/L EDTA·4Na
Storage & Handling
Storage: -30~-5 ℃, protected from light. Shelf life: 24 months. Ship cold-chain where required.
Quality & Documentation
Manufactured under ISO-aligned quality management systems. Full documentation — Certificate of Analysis (CoA), Material Safety Data Sheet (MSDS), formulation summary, and stability data — is available on request. Email [email protected] with the SKU.
About this product
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Learn more about this
- Counting Cells with a HemocytometerA hemocytometer is a thick glass slide bearing a precisely etched grid over which a coverslip creates a chamber exactly 0.1 mm deep, so that a known volume of cell suspension can be counted under a microscope and converted to a concentration. For mammalian cells you count the four 1 mm2 corner squares, take the mean, multiply by the dilution factor, and multiply by 10^4 to obtain cells per mL, because each corner square encloses exactly 10^-4 mL. Mixing the sample 1:1 with 0.4% trypan blue before loading gives a viability percentage from the same count, since only dead cells with compromised membranes take up the dye.
- Trypsin-EDTATrypsin-EDTA is a cell dissociation reagent that combines the serine protease trypsin, which cleaves peptide bonds on the C-terminal side of lysine and arginine residues in cell-surface and matrix proteins, with the chelator EDTA, which binds the calcium and magnesium ions that cell-adhesion molecules require. It is supplied in a calcium- and magnesium-free balanced salt solution, most commonly at 0.05% trypsin (0.5 g/L) for routine cell lines and 0.25% (2.5 g/L) for firmly adherent cells and primary cultures. Typical use is 2-5 minutes at 37 degrees C, followed immediately by neutralisation with serum-containing medium or a defined trypsin inhibitor.
- Subculture of Cells (Passaging)Subculture of cells, also called passaging, is the transfer of cells from a culture that is approaching confluence into fresh vessels with fresh medium so that growth can continue. Adherent cells are detached first, usually with 0.25% or 0.05% trypsin-EDTA or a non-enzymatic dissociation reagent, then reseeded at a lower density set by a split ratio such as 1:4; suspension cells are simply diluted into fresh medium without any dissociation step. Most continuous adherent cell lines are subcultured at 70-80% confluence, which normally means two or three passages per week.
- Trypsin vs Accutase: Choosing a Cell Dissociation ReagentTrypsin is a pancreatic serine protease that cleaves peptide bonds after lysine and arginine, and it detaches cells fast but indiscriminately - it must be stopped with serum or a trypsin inhibitor, and it cleaves surface proteins along with the adhesion contacts. Accutase is a proprietary mixture of proteolytic and collagenolytic enzymes from an invertebrate source, supplied in calcium- and magnesium-free DPBS with 0.5 mM EDTA; it works more slowly and gently, does not require a neutralisation step, and preserves more surface epitopes. Choose trypsin for routine passaging of robust adherent lines where speed matters and the cells go straight back into culture; choose Accutase when the harvested cells are the experiment - flow cytometry on surface markers, pluripotent stem cells, primary neurons, or any single-cell suspension where viability and phenotype must survive the harvest.
- DispaseDispase, also sold as Dispase II or neutral protease, is a zinc-dependent metalloprotease from Paenibacillus (formerly Bacillus) polymyxa, EC 3.4.24.4, used for gentle tissue dissociation and for detaching cells and epithelial sheets intact. It cleaves fibronectin and type IV collagen, degrades type I collagen only minimally, and does not cleave laminin or type V collagen, which is why it can separate an epidermis from a dermis or lift a confluent epithelial sheet off plastic without destroying cell-cell junctions. Because it is a metalloprotease requiring zinc for catalysis and calcium for stability, it is inhibited by EDTA, EGTA and 1,10-phenanthroline, and it must be diluted in a calcium-containing buffer rather than a chelator-based one.
