SignalSilence siRNA
SignalSilence® Atg4B siRNA I #6336
| Item # | Description | List Price | Web Price | QTY | Buy |
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| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
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| H |
Applications Key:
Reactivity Key:
H = Human
Species cross-reactivity is determined by Western blot.
Description
SignalSilence® Atg4B siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Atg4B 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 blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® Atg4B siRNA I (+), using Atg4B Antibody #5299 (upper) or β-Tubulin (9F3) Rabbit mAb #2128 (lower). The Atg4B Antibody confirms silencing of Atg4B expression, while the β-Tubulin (9F3) Rabbit mAb is used as a loading control.
Directions for Use
CST recommends transfection with 100 nM Atg4B 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.
Background
Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents. Control of autophagy was largely discovered in yeast and involves proteins encoded by a set of autophagy-related genes (Atg) (1). Formation of autophagic vesicles requires a pair of essential ubiquitin-like conjugation systems, Atg12-Atg5 and Atg8-phosphatidylethanolamine (Atg8-PE), which are widely conserved in eukaryotes (2). Numerous mammalian counterparts to yeast Atg proteins have been described, including three Atg8 proteins (GATE-16, GABARAP, and LC3) and four Atg4 homologues (Atg4A/autophagin-2, Atg4B/autophagin-1, Atg4C/autophagin-3, and Atg4D/autophagin-4) (3-5). The cysteine protease Atg4 is pivotal to autophagosome membrane generation and regulation. Atg4 primes the Atg8 homologue for lipidation by cleaving its carboxy terminus and exposing its glycine residue for E1-like enzyme Atg7. The Atg8 homologue is transferred to the E2-like enzyme Atg3 before forming the Atg8-PE conjugate. During later stages of autophagy, Atg4 can reverse this lipidation event by cleaving PE and recycle the Atg8 homologue (6).
- Reggiori, F. and Klionsky, D.J. et al. (2002) Eukaryot Cell1, 11 - 21.
- Ohsumi, Y. et al. (2001) Nat Rev Mol Cell Biol2, 211 - 6.
- Kabeya, Y. et al. (2000) EMBO J19, 5720 - 8.
- Kabeya, Y. et al. (2004) J Cell Sci117, 2805 - 12.
- Mariño, G. et al. (2003) J Biol Chem278, 3671 - 8.
- Sou, Y.S. et al. (2008) Mol Biol Cell19, 4762 - 75.
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
- 6201 SignalSilence® Control siRNA (Fluorescein Conjugate)
- 6568 SignalSilence® Control siRNA (Unconjugated)
- 5299 Atg4B Antibody
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
