TranslationBlocker Human ApoE siRNA – DISCONTINUED

DISCONTINUED: Human ApoE TranslationBlocker™ siRNA Duplex Product Attributes

ApoE Knockdown Protocol:

1. 1. Starting with cultured cells in growth media seed 24 well plates with 5 x 104 cells and incubate at 37°C for 24 hours.
   2. Prepare transfection reagent in serum-free growth media allow to come to room temperature. The above data was generated with Oligofectamine prepared in accordance with manufacturer’s instructions.
   3. Dilute siRNA to working concentration in serum-free growth media. Optimal dilution should be experimentally determined.
   4. Combine diluted siRNA and transfection reagent. Equivalent volumes of prepared Oligofectamine and diluted siRNA should be combined and incubated at room temperature 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 without a media change. Generally 1-2 fold of the original volume of siRNA containing serum free media is added to restore nutrient levels in media.
   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, Apolipoprotein E (ApoE) knockdown at the protein level was not visible until 24 hours post transfection. Knockdown continued to improve