TranslationBlocker Human Alpha-synuclein siRNA – DISCONTINUED

DISCONTINUED: Human Alpha-synuclein TranslationBlocker™ siRNA Duplex Product Attributes

1. 1. Starting with cultured cells in growth media (+10% fetal calf serum (FCS)) seed 6 well plates with 3 x 105 cells and incubate at 37°C for 24 hours.
   2. Prepare transfection reagent in serum-free growth media (Opti-MEM used in this experiment) and allow to come to room temperature. The above data was generated with Lipofectamine 2000 diluted 1:100 prepared in accordance with manufacturer’s instructions.
   3. Dilute siRNA to working concentration in serum-free growth media. The above data was generated with 60-120 pmol of QX2 siRNA duplex in 300 ul of media. Optimal dilution should be experimentally determined.
   4. Combine diluted siRNA and transfection reagent. In this case, 100 ul of Lipofectamine 2000 was incubated with 100 ul of diluted siRNA for 30 minutes with gentle mixing (optional).
   5. Add prepared siRNA solution to cells.
   6. Incubate for several hours in serum free media. In the above experiment, a 6 hour incubation was performed.
   7. Add serum containing media to culture. Data above was generated by adding 1.5x culture volume of media containing 30% FBS (2% final concentration).
   8. Incubate and assay several hours after transfection. An additional transfection may be performed, as above, 24 hours after initial transfection to enhance knockdown. Optimal knockdown time should also be experimentally determined. In the above experimental conditions, SNCA knockdown at the protein level was confirmed 72 hours post transfection via western blot.