0.25% Trypsin-EDTA (1×)

• Ready to Use: No additional dilution or preparation required; can be directly used for cell dissociation, saving experimental setup time.

• Strong & Fast Dissociation: 0.25% trypsin efficiently dissociates tightly adherent cell clusters, significantly reducing digestion time.

• Synergistic Action with EDTA: EDTA chelatesCa2+ and Mg2+, enhancing the disruption of cell-cell junctions and improving dissociation uniformity.

• Consistent Quality: Manufactured with high-purity trypsin, sterile-filtered to ensure cell culture safety and reproducibility.

• Broad Compatibility: Suitable for routine passaging and dissociation of a wide range of mammalian cell lines.

For dissociation, passaging, and collection of adherent cells.

1. Preparation

a) Take the trypsin solution from the refrigerator and pre-warm it in a 37 ℃ water bath or incubator for 5-10 minutes.

Note: Do not prewarm the entire bottle; only take out the volume you need.

b) Remove the old medium from the culture dish or flask, and gently rinse once with sterile PBS, Hanks, or serum-free medium to remove residual serum.

Note: Serum inhibits trypsin activity.

2. Trypsinization

a) Add an appropriate amount of trypsin solution to just cover the cell layer. Incubate at room temperature for 1-5 minutes, gently swirling the culture vessel occasionally to ensure even contact of trypsin with the cells.

b) Observe under a microscope. dissociation is sufficient when cells begin to round up and detach from the surface.

3. Termination of Trypsinization

a) Immediately add an equal volume or twice the volume of complete medium containing serum to stop the action of trypsin.

b) Gently pipette to mix, ensuring the cells are fully detached into a single-cell suspension.

4. Cell Collection and Culture

a) Transfer the cell suspension to a centrifuge tube and centrifuge at 1,000 rpm for 3-5 minutes.

b) Discard the supernatant and resuspend the cells in fresh culture medium. Proceed with counting, passaging, or seeding according to experimental requirements.

Store at 4 ℃ for 3 months; -20 ℃ for 2 years.

1.After use, please immediately seal and store at 4 ℃. For long-term storage, keep at -20 ℃ and avoid repeated freeze-thaw cycles.

2.Avoid prolonged trypsin dissociation, as this may cause cell damage or death.

3.For sensitive cells (e.g., primary cells or stem cells), consider shortening the dissociation time or diluting the trypsin concentration.

4.Observe aseptic techniques to prevent microbial contamination.

5.The product is for R&D use only, not for diagnostic procedures, food, drug, household or other uses.

6.This product may irritate skin, eyes, and the respiratory tract. Please wear a lab coat and disposable gloves.

• C0202-Instruction Manual(206 KB)