Cell Signaling Technology

Product Pathways - Apoptosis

SignalSilence® HtrA2/Omi siRNA I #7574

Applications Reactivity
Transfection H (Mk)

Reactivity Key:  H=Human  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® HtrA2/Omi siRNA I (+), using HtrA2/Omi (D20A5) Rabbit mAb #9745 (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The HtrA2/Omi (D20A5) Rabbit mAb confirms silencing of HtrA2/Omi expression, while the GAPDH (D16H11) XP® Rabbit mAb is used as a loading control.

Description

SignalSilence® HtrA2/Omi siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit HtrA2/Omi expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.

Quality Control

Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.

Directions for Use

CST recommends transfection with 100 nM SignalSilence® HtrA2/Omi siRNA I 48 to 72 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.

Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.

Background

High temperature requirement protein A2 (HtrA2)/Omi is a serine protease with homology to the E. coli HtrA protein (DegP) and is thought to be involved in apoptosis and stress-induced degradation of misfolded proteins (1). While HtrA2 was orignally identified to be present in either the nucleus (1) or endoplasmic reticulum (2), subsequent studies have shown that it localizes in mitochondria and is released during apoptosis (3-8). HtrA2 is produced as a 50 kDa zymogen that is cleaved to generate a 36 kDa mature protein that exposes an amino terminal motif (AVPS) resembling that of the IAP inhibitor Smac/Diablo (3-8). Like Smac, interaction between HtrA2 and IAP family members, such as XIAP, antagonizes their inhibition of caspase activity and protection from apoptosis (3-8). Interestingly, HtrA2 knock-out mice did not show signs of reduced apoptosis, but rather had a loss of neurons in the striatum and a Parkinson's-like phenotype, suggesting that HtrA2 might have a neuroprotective function (9-11). This activity is associated with the protease activity of HtrA2 (9). Furthermore, research studies have shown that loss of function mutations in the HtrA2 gene are associated with Parkinson's disease (12).

  1. Gray, C.W. et al. (2000) Eur. J. Biochem. 267, 5699-5710.
  2. Faccio, L. et al. (2000) J. Biol. Chem. 275, 2581-2588.
  3. Suzuki, Y. et al. (2001) Mol. Cell 8, 613-621.
  4. Hegde, R. et al. (2002) J. Biol. Chem. 277, 432-438.
  5. Martins, L.M. et al. (2002) J. Biol. Chem. 277, 439-444.
  6. van Loo, G. et al. (2002) Cell Death Differ. 9, 20-26.
  7. Verhagen, A.M. et al. (2002) J. Biol. Chem. 277, 445-454.
  8. Martins, L.M. et al. (2002) J. Biol. Chem. 277, 439-444.
  9. Jones, J.M. et al. (2003) Nature 425, 721-727.
  10. Vaux, D.L. and Silke, J. (2003) Cell 115, 251-253.
  11. Martins, L.M. et al. (2004) Mol. Cell Biol. 24, 9848-9862.
  12. Strauss, K.M. et al. (2005) Hum. Mol. Genet. 14, 2099-2111.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

Limited Use Label License, RNA interference: This product is licensed under European Patent 1144623 and foreign equivalents from Ribopharma AG, Kulmbach, Germany and is provided only for use in non-commercial research specifically excluding use (a) in drug discovery or drug development, including target identification or target validation, by or on behalf of a commercial entity, (b) for contract research or commercial screening services, (c) for the production or manufacture of siRNA-related products for sale, or (d) for the generation of commercial databases for sale to Third Parties. Information about licenses for these and other commercial uses is available from Ribopharma AG, Fritz-Hornschuch-Str. 9, D-95326 Kulmbach, Germany.

For Research Use Only. Not For Use In Diagnostic Procedures.

Products