Product Pathways - NF-kB Signaling
SignalSilence® NF-κB p65 siRNA I #6261
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Western blot analysis of extracts from HeLa cells transfected with nontargeted (-) or targeted (+) siRNA. p65 was detect using NF-kappaB p65 Antibody #3034, and p42 was detected using p42 MAPK Antibody #9108. The NF-kappaB p65 Antibody confirms silencing of p65 expression, and the p42 MAPK Antibody is used to control for loading and siRNA specificity.
SignalSilence® NF-kappaB p65 siRNA from Cell Signaling Technology allows the researcher to specifically inhibit NF-kappaB p65 expression using RNA interference, a method in which gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products are rigorously tested in-house and have been shown to reduce protein expression.
Directions for Use
CST recommends transfection with 100 nM NF-kappaB p65 siRNA 48 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.
Transcription factors of the nuclear factor κ B (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, c-Rel, RelB, NF-κB1 (p105/p50), and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. Rel proteins bind p50 and p52 to form dimeric complexes that bind DNA and regulate transcription. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins (3-5). NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKKα (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which translocates to the nucleus (9-11).
Small Interfering RNA (siRNA) has been used to specifically silence NF-kappaB p65 in colon cancer cells (10).
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- 6568 SignalSilence® Control siRNA (Unconjugated)
- 6201 SignalSilence® Control siRNA (Fluorescein Conjugate)
- 2694 Phospho-IKKα/β (Ser176/180) Antibody II
- 2695 IKKγ (DA10-12) Mouse mAb
- 3033 Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb
- 3036 Phospho-NF-κB p65 (Ser536) (7F1) Mouse mAb
- 4954 RelB Antibody
- 3039 Phospho-NF-κB p65 (Ser468) Antibody
- 4812 IκBα (44D4) Rabbit mAb
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
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.