TranslationBlocker Mouse Apoptosis regulator BAX siRNA – DISCONTINUED

DISCONTINUED: Mouse Apoptosis regulator BAX TranslationBlocker™ siRNA Duplex Product Attributes

1. 1. Starting with cultured cells in growth media seed 6 well plates with cells and incubate at 37°C until cells reach 40% confluence. In the example data, larger cultures of 2 x 10⁵ were transfected
   2. Prepare transfection reagent in serum-free growth media such as Opti-MEM allow to come to room temperature. The above data was generated with Oligofectamine for delivery of mouse BAX siRNA prepared in accordance with manufacturer’s instructions.
   3. Dilute siRNA to working concentration in serum-free growth media. For Oligofectamine or similar transfections into 6 well plates, we recommend starting siRNA concentration optimization at around 150 pmol siRNA duplex in 300 ul of serum free media. Optimal dilution should be experimentally determined.
   4. Combine diluted siRNA and transfection reagent. Equivalent volumes of prepared Oligofectamine or other transfection reagent 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. Additionally, FITC labeled control siRNA (Product Number QC-1F) or positive control siRNA (Beta-Actin; Product Number QC-2) may be used to assess transfection efficiency.
   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. Optimal knockdown time should also be experimentally determined. Optionally, a second transfection can be performed 24 hours after the first transfection to enhance knockdown. Mouse BAX knockdown at the protein level was measured at 48 hours post transfection via western blot.