Cell Signaling Technology

Product Pathways - Stem Cell and Lineage Markers

SignalSilence® Sall4 siRNA II #12111

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 NTERA-2 cl.D1 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® Sall4 siRNA I #12136 (+), or SignalSilence® Sall4 siRNA II (+), using Sall4 (D16H12) Rabbit mAb #8459 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The Sall4 (D16H12) Rabbit mAb confirms silencing of Sall4 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

Description

SignalSilence® Sall4 siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Sall4 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® Sall4 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

Members of the SALL gene family encode putative zinc finger transcription factors highly expressed during development (1). Sall4 is expressed very early in development with other pluripotency regulators, such as Oct-4 and Nanog (2). Recent studies suggest Sall4 works as a master regulator that controls its own expression and the expression of Oct-4 in a transcriptional regulation feedback loop governing stem cell pluripotency and stem cell fate (2,3). Immunohistochemical studies indicate that Sall4 is a sensitive and specific diagnostic marker for primary germ cell tumors and yolk sac tumors (4,5). Research studies have shown that Sall4 is constitutively expressed in acute myeloid leukemia (AML) and is a probable effector of the Wnt/β-catenin signaling pathway in this disease (6). In addition, mutations in Sall4 have been implicated in human malformation syndromes including Duane-radial ray syndrome (Okihiro syndrome) and Acro-renal-ocular syndrome (7).

  1. Sweetman, D. and Münsterberg, A. (2006) Dev Biol 293, 285-93.
  2. Yang, J. et al. (2008) Proc Natl Acad Sci USA 105, 19756-61.
  3. Yang, J. et al. (2010) PLoS One 5, e10766.
  4. Mei, K. et al. (2009) Mod Pathol 22, 1628-36.
  5. Miertus, J. et al. (2006) Hum Genet 119, 154-61.
  6. Ma, Y. et al. (2006) Blood 108, 2726-35.
  7. Al-Baradie, R. et al. (2002) Am J Hum Genet 71, 1195-9.

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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