| Field | Specification |
|---|---|
| Species |
- Protein expression: p53 positive
- Isoenzymes: PGM3, 1, PGM1, 1, ES-D, 1, AK-1, 1-2, GLO-1, 2, G6PD, B
- Tumorigenic: Yes, in nude mice. Forms malignant melanoma (large round cell type)
- Products: Melanin
- Mutational profile: BRAF V600E mut: V600E type BRAF Mutation was determined by DNA based methods (sequencing, RT-PCR) and protein based methods (Western Blot), N-Ras wt
- cultureMedium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: Leave at least 48 hours post to thawing until removal of medium or subculture
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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