Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

SignalSilence® HELLS siRNA II #11823

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 DLD-1 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® HELLS siRNA I #11822 (+), or SignalSilence® HELLS siRNA II (+), using HELLS Antibody #7998 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The HELLS Antibody confirms silencing of HELLS expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

Description

SignalSilence® HELLS siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit HELLS 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® HELLS siRNA II 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

HELLS, or LSH1, is a lymphoid-specific helicase thought to be involved in cellular proliferation and leukemogenesis (1,2). It is believed to be a chromatin remodeler and is required for DNMT1-mediated methylation maintenance and DNMT3A/DNMT3B-mediated de novo methylation. The role of HELLS in methylation maintenance was thought to be largely confined to repetitive DNA sequences, including major and minor satellite sequences, rather than single copy genes (3,4); recent evidence suggests a role in genome-wide cysteine methylation at non-repeat sequences (5). De novo methylation maintenance is associated with silencing of specific genes, some known to be involved in pluripotency and lineage commitment (6,7).

  1. Lee, D.W. et al. (2000) Cancer Res 60, 3612-22.
  2. Fan, T. et al. Epigenetics 3, 134-42.
  3. Dennis, K. et al. (2001) Genes Dev 15, 2940-4.
  4. Muegge, K. (2005) Biochem Cell Biol 83, 548-54.
  5. Tao, Y. et al. (2011) Proc Natl Acad Sci USA 108, 5626-31.
  6. Xi, S. et al. (2009) Stem Cells 27, 2691-702.
  7. Xi, S. et al. (2007) Proc Natl Acad Sci USA 104, 14366-71.

Application References

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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.

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