TranslationBlocker Human Apoptosis regulator BAX siRNA

Human Apoptosis regulator BAX TranslationBlocker™ siRNA Duplex Product Attributes

Apoptosis regulator BAX Knockdown Protocol:

1. 1. Starting with cultured cells in growth media seed 6 well plates with 5 x 104 cells and incubate at 37°C until cells reach 40% confluence
   2. Prepare transfection reagent in serum-free growth media allow to come to room temperature. The above data was generated with 8 ul of Oligofectamine prepared in accordance with manufacturer’s instructions.
   3. Dilute siRNA to working concentration in serum-free growth media. The above data used 200 pmol siRNA duplex. 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. Optionally, as with the above data, a second transfection can be performed 24 hours after the first transfection to enhance knockdown. BAX knockdown at the protein level was measured at 48 hours post transfection via western blot.