TranslationBlocker Human Amyloid beta A4 protein siRNA

Human Amyloid beta A4 protein TranslationBlocker™ siRNA Duplex Product Attributes

Amyloid beta A4 protein Knockdown Protocol:

1. 1. Starting with cultured cells in growth media 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 I used in this experiment) and allow to come to room temperature. The above data was generated with Lipofectamine 2000 diluted 1:25 prepared in accordance with manufacturer’s instructions.
   3. Dilute siRNA to working concentration in serum-free growth media. The above data was generated with 100 pmol of QX3 siRNA duplex in 250 ul of media. Optimal dilution should be experimentally determined.
   4. Combine diluted siRNA and transfection reagent. In this case, 250 ul of Lipofectamine 2000 was incubated with 250 ul of diluted siRNA for 20 minutes with gentle mixing (optional).
   5. Add prepared siRNA solution to cells.
   6. Incubate for several hours in serum free media.
   7. Add serum containing media to culture.
   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, Amyloid Beta A4 knockdown at the protein level was confirmed 48 hours post transfection via western blot.