This non-enzymatic cell detachment solution contains no proteolytic enzymes, yet effectively detaches adherent cells from the surface of culture flasks and dishes for cell isolation. Compared with conventional trypsin digestion, it acts mildly without damaging or altering cellular biological properties, making it the optimal cell dissociation reagent for in vivo tumor cell transplantation assays.After detachment with this solution, cells can be directly subcultured or harvested via centrifugation for downstream experiments. Free of protease additives, it is ideal for cell collection in nuclear & cytoplasmic protein extraction, Western Blot and co-immunoprecipitation assays. It prevents protein degradation in samples and avoids RNA breakdown induced by RNase contamination present in protease preparations.
Product Features
1. Adherent cells fully detach from culture vessels after 10 minutes of incubation at room temperature, delivering gentle yet efficient dissociation.
2. Dissociated cells support continuous serial subculture.
3. The preferred dissociation method for tumor cell culture prior to in vivo transplantation studies.
4. Eliminates excessive proteolytic damage to cells and sample protein degradation.
5. Suitable for nuclear/cytoplasmic protein extraction, Western Blot and co-immunoprecipitation.
6. Prevents RNA degradation caused by RNase contaminants in protease reagents.
7. Formulated with EDTA; trypsin-free.
Instructions for Use
1. Rinse the cell monolayer with PBS and aspirate all residual liquid thoroughly from the vessel.
2. Add an adequate volume of non-enzymatic cell detachment solution to fully cover the cell layer. Rock the flask/dish gently to ensure uniform contact between all cells and the reagent.
3. Incubate at room temperature for 10 minutes (or adjusted duration), rocking the vessel occasionally until complete cell detachment. Quench the reaction by adding complete culture medium or 2–3 volumes of PBS buffer. Clustered cell aggregates after detachment are normal; such aggregates actually promote tumor cell growth for transplantation applications. Dissociation proceeds faster at 37 °C.
4. Centrifuge the cell suspension at 500 g for 5–10 minutes, discard the supernatant and collect the cell pellet.
5. Harvested cells can be applied to the following experiments:
(1) Nuclear and cytoplasmic protein extraction
(2) Western Blot analysis
(3) Co-immunoprecipitation
(4) DNA-protein binding assays
(5) RNA isolation
(6) Cell splitting and subculture
Precautions
1. This product must not be freeze-thawed to prevent inactivation.
2. Strictly avoid bacterial contamination of the detachment solution during operation.
3. Do not over-digest cells; prolonged incubation will compromise post-seeding cell viability and growth.
4. Wear a lab coat and disposable gloves during manipulation for personal safety.
