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| Product Name | Dispase II |
| Catalog No. | RESK-0238 |
| Description | Dispase II is a non-specific neutral metalloprotease commonly used in cell biology to isolate single cells from various tissues and organs for subsequent cell culture, such as primary cell isolation and cell passaging. It can also be used to prevent or eliminate cell aggregation that occurs during suspension cell culture. This product is supplied as a non-sterile lyophilized powder derived from Bacillus polymyxa and must be sterilized (e.g., by filtration) before use. For cell culture applications, the typical working concentration is 0.6-2.4 U/mL. Dispase II is rapid, effective, and gentle, causing minimal damage to cells while maintaining cell membrane integrity. |
| Intended Use | For research use in the dissociation of various tissue types, isolation of primary cells, cell passaging, organoid isolation, and prevention/elimination of cell aggregation in suspension cultures. |
| Principle / Technology | Dispase II is a neutral, non-specific metalloprotease that gently cleaves extracellular matrix and intercellular junction proteins, releasing single cells from tissues with minimal damage while maintaining cell membrane integrity. Compared with other commonly used proteases (e.g., trypsin, collagenase, pronase), it is rapid and gentle, of bacterial origin (free of mycoplasma and animal virus contamination), highly stable, and unaffected by temperature, pH, or serum components. |
| Sample Type | Tissues and organs of various types; adherent and suspension cultured cells; primary cells; organoids |
| Performance Range / Specifications | Non-sterile lyophilized powder; requires sterilization before use Recommended working concentration for cell separation: 0.6-2.4 U/mL Rapid, effective, and gentle; maintains cell membrane integrity Bacterial origin; free of mycoplasma and other animal virus contamination High stability; not affected by temperature, pH, or serum components |
| Specificity | Non-specific neutral metalloprotease |
| Reaction Conditions / Protocol | 1. Preparation of stock and working solutions 1) Dissolve the lyophilized powder in 50 mM HEPES-KOH buffer (pH 7.4, 150 mM NaCl) to prepare a 10 mg/mL stock solution; sterilize by filtration through a 0.22 um filter. 2) Dilute the stock solution with appropriate cell culture medium to the working concentration before use. For cell separation, the typical working concentration is 0.6-2.4 U/mL. Note: Working concentrations above 2.4 U/mL are not recommended. 2. Tissue dissociation 1) Mince tissue into appropriately sized pieces with a sterile scalpel or scissors. 2) Wash the tissue pieces with sterile PBS. 3) Add Dispase II solution (0.6-2.4 U/mL), ensuring the tissue is fully immersed. 4) Incubate at 37 °C with slow agitation until fully dissociated. For difficult tissues, 1 hour is generally sufficient; longer incubation (several hours) does not significantly affect cell viability. For first use, determine total incubation time by cell counting. 5) If needed, pass the digest through a sterile stainless steel mesh to separate single cells from residual tissue; alternatively, let large fragments settle and gently decant the supernatant cells. Fresh enzyme solution may be added for further dissociation of residual tissue. 6) Pellet the cells by centrifugation and discard the enzyme solution. 7) Resuspend the cell pellet in culture medium and culture under routine conditions. 3. Cell passaging 1) Cover the cells with Dispase II solution (pre-warmed to 37 °C) and incubate at 37 °C for 5 min. 2) Aspirate the solution and continue incubation at 37 °C for 10 min. 3) Observe cell detachment under a microscope; incubate for an additional 15 min if needed. 4) Suspend the cells in culture medium, gently spin down, and wash with medium. 5) Resuspend in fresh medium and plate according to routine methods. |
| Components / Formulation | Non-sterile lyophilized Dispase II powder (neutral metalloprotease from Bacillus polymyxa). |
| Storage Conditions | Store the lyophilized powder at 4 °C (shelf life 2 years). The reconstituted stock solution is stable for 2 weeks at 4 °C; for long-term storage, aliquot and freeze at -20 °C. Avoid repeated freeze-thaw cycles. |
| Shelf Life | 2 years (lyophilized powder at 4 °C); stock solution stable for 2 weeks at 4 °C. |
| Package Specifications | 100 mg 1 g |
| Product Form | Lyophilized powder (non-sterile) |
| Quality Control | Free of mycoplasma and other animal virus contamination (bacterial origin). |
| Concentration | Stock solution: 10 mg/mL; recommended working concentration: 0.6-2.4 U/mL |
| Activity / Unit Definition | Recommended working concentration for cell separation: 0.6-2.4 U/mL (do not exceed 2.4 U/mL). Specific activity per mg is not specified. |
| Source / Origin | Bacillus polymyxa (bacterial) |
| pH Range / Optimal pH | Neutral protease; reconstitution buffer at pH 7.4 is recommended. Activity is stable and not affected by pH within typical culture conditions. |
| Expiration Date / Stability | Lyophilized powder: 2 years at 4 °C. Stock solution: 2 weeks at 4 °C; for long-term storage, aliquot and store at -20 °C; avoid repeated freeze-thaw cycles. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Suitable for separation of many tissue and cell types, primary cells, and organoids; activity is not affected by serum components. |
| Recommended Buffer System | Dissolve in 50 mM HEPES-KOH buffer (pH 7.4, 150 mM NaCl). Use of the recommended HEPES buffer is advised; other buffers have not been validated. |
| Application Notes / Precautions | This product is non-sterile and must be filter-sterilized (0.22 um) before use. Do not use working concentrations above 2.4 U/mL. When using for the first time, determine the optimal total incubation time by cell counting. Prolonged incubation (several hours) does not significantly affect cell viability. |
For research use only, not for clinical use.
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