Cell Signaling Technology

Product Pathways - Metabolism

SignalSilence® OPA1 siRNA I #67677

No. Size Price
67677S 300 µl ( 3 nmol ) ¥3,224.00 现货查询 购买询价
67677 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
TFN Human,

Species cross-reactivity is determined by western blot.

Applications Key: TFN=Transfection,

Description

SignalSilence® OPA1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit OPA1 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.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® OPA1 siRNA I (+), or SignalSilence® OPA1 siRNA II #89128 (+) using OPA1 (D7C1A) Rabbit mAb #67589 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The OPA1 (D7C1A) Rabbit mAb confirms silencing of OPA1 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

Directions for Use

CST recommends transfection with 100 nM SignalSilence® OPA1 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

Changes in mitochondrial dynamics regulated by environmental cues affect mitochondrial size and shape and have been shown to dramatically impact mitochondrial metabolism, apoptosis, and autophagy (1). These processes are largely controlled by mitochondrial dynamin-related GTPases, including mitofusin-1, mitofusin-2, OPA1, and DRP1. DRP1 regulates mitochondrial fission, while the mitofusins and OPA1 control fusion at the outer and inner mitochondrial membrane, respectively.

OPA1, or Optic Atrophy 1, was originally identified as a genetic cause for Autosomal Dominant Optic Atrophy, a neuropathy resulting in progressive visual loss (2,3). OPA1 is a widely expressed protein localized to the inner mitochondrial membrane, which regulates mitochondrial fusion and cristae morphology and protects against apoptosis (4-6). OPA1 activity is tightly regulated through alternative splicing and post-translational modifications including complex proteolytic processing by multiple proteases (7-12). In addition, OPA1 expression can be induced under conditions of metabolic demand through a pathway involving Parkin induced NF-κB activation (13).

  1. Kasahara, A. and Scorrano, L. (2014) Trends Cell Biol 24, 761-70.
  2. Delettre, C. et al. (2000) Nat Genet 26, 207-10.
  3. Alexander, C. et al. (2000) Nat Genet 26, 211-5.
  4. Frezza, C. et al. (2006) Cell 126, 177-89.
  5. Olichon, A. et al. (2003) J Biol Chem 278, 7743-6.
  6. Griparic, L. et al. (2004) J Biol Chem 279, 18792-8.
  7. Delettre, C. et al. (2001) Hum Genet 109, 584-91.
  8. Olichon, A. et al. (2007) Cell Death Differ 14, 682-92.
  9. Ishihara, N. et al. (2006) EMBO J 25, 2966-77.
  10. Cipolat, S. et al. (2006) Cell 126, 163-75.
  11. Griparic, L. et al. (2007) J Cell Biol 178, 757-64.
  12. Merkwirth, C. et al. (2008) Genes Dev 22, 476-88.
  13. Müller-Rischart, A.K. et al. (2013) Mol Cell 49, 908-21.

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!

Protocols

Companion Products


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

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

SignalSilence is a registered trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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