| Field | Specification |
|---|---|
| Mfr No | |
| Species |
- Receptors expressed: Epidermal growth factor (EGF), multiplication stimulating activity (MSA)
- Protein expression: IBB-DiHcRed1: Location/gene: 1..589 / Pcmv, 656..916 / IBB, 932..1615 , 1670..2356
- / HcRed1, 3587..4381 / KanR/NeoR
- Products: CMV Promotor IBB (Ribbeck & Gorlich 2002), Neomycin, Phosphotransferase, Epidermal growth factor, multiplication stimulating activity
- 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, 0.5 mg/mL G418
- dissociationReagent: Accutase
- subculturing: Discard the old medium and wash the cells with PBS. Add a freshly prepared 0.025% trypsin/0.02% EDTA solution heated to 37 degrees Celsius and wait until the cells detach, which usually takes about 5 minutes. Neutralize the trypsin by adding fresh medium, then transfer the cell mixture to a tube and centrifuge. After centrifugation, remove the supernatant, resuspend the cell pellet in fresh culture medium, and transfer the suspension to new flasks. Incorporate G418 into the culture medium to achieve a final concentration of 0.5 mg/ml
- seedingDensity: 2 to 4 x 104 cells/cm2
- fluidRenewal: 2 to 3 times per week
- 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.