Tools for siRNA Western Blotting

Thermo Scientific Pierce Validated Intracellular Antibodies are cellular proteomics antibodies that have been thoroughly tested ensure specific Western blot detection of target proteins in siRNA experiments. When combined with the appropriate siRNA reagents and Western blot detection reagents, each validated antibody enables accurate measurement of the changes in protein expression caused by gene silencing.

Looking for the siRNA Western Blot Kits?

Previously, we offered complete Thermo Scientific SuperSignal siRNA Western Blot Kits corresponding to each of the validated antibodies. Each kit included all the general and specific components needed for both the siRNA experiment and the Western blot detection step using the specific antibody. Although we no longer offer these complete kits because of the costs associated with maintaining so many specialized kit assemblies, we continue to offer all of the components needed to perform the technique. Offering the components separately provides you with greater flexibility to purchase the amounts and combinations of reagents needed for your customized experimental needs.

Essential products for siRNA Western blotting:

The following Thermo Scientific Products are needed to perform the siRNA-Western blotting technique:

• Thermo Scientific Dharmacon siRNA Reagents:
  • ON-TARGETplus SMARTpool siRNA for the specific gene of interest
  • ON-TARGETplus siCONTROL Non-Targeting Pool
  • siGLO Green Transfection Indicator
  • 5X siRNA Buffer
     
• Pierce Validated Antibody:
  • Select the antibody specific for the protein product of the target gene
     
• Pierce Western Blotting Reagents:
  • Select individual Western blotting reagents: choose and optimize your experiment with an appropriate blocking buffer (e.g., StartingBlock Buffer, Part No. 37539), wash buffer (e.g., Part No. 28352), HRP-conjugated secondary antibody (anti-Mouse IgG or anti-Rabbit IgG, depending on species of validated antibody used), and SuperSignal Chemiluminescent Substrate ("Dura" formulation recommended, Part No. 34075)

Posters describing siRNA Western blotting:

1. Webb, B.L., et al. (2007). Analysis of signaling pathways involved in TNFa-mediated cytokine production in A549 cells using RNA interference and antibody microarrays.
2. Narahari, J., et al. (2007). Validation of siRNA-mediated gene silencing using downstream functional analysis.
3. Webb, B.L, et al. (2007). Analysis of signaling pathways using antibody microarrays and RNA interference.
4. Hughes, D.E., Hong, S.J. and Webb, B.L. (2007). Cell-based screening applications utilizing cryo-preserved siRNA-transfected cells.