TranslationBlocker Human Androgen receptor siRNA – DISCONTINUED

DISCONTINUED: Human Androgen receptor TranslationBlocker™ siRNA Duplex Product Attributes

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 such as Opti-MEM. The above data was generated with 100 pmol of QX4 siRNA duplex in 250 ul of 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, Androgen Receptor knockdown at the protein level was confirmed 48 hours post transfection via western blot.