Cell Signaling Technology

Product Pathways - DNA Damage

SignalSilence® Miz-1 siRNA I #12335

Applications Reactivity
Transfection H (M) (R) (Mk)

Reactivity Key:  H=Human  M=Mouse  R=Rat  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 HaCaT cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® Miz-1 siRNA I (+), or SignalSilence® Miz-1 siRNA II #12405 (+), using Miz-1 Antibody #9044 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The Miz-1 Antibody confirms silencing of Miz-1 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

Description

SignalSilence® Miz-1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Miz-1 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® Miz-1 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

Miz-1 (Zbtb17) is a poxvirus and zinc finger (POZ) transcription factor with an amino-terminal BTB/POZ domain and 13 carboxy-terminal zinc finger domains. Miz-1 plays a key role in cell cycle control through activation of the cyclin-dependent kinase inhibitors p15, INK4B, and p21 Waf1/Cip1 (1-4). The transcriptional activity of Miz-1 is repressed through direct interaction with Myc (1-4). In the presence of DNA damage, Myc is recruited to the p21 Waf1/Cip1 promoter by Miz-1 and blocks p53-mediated induction of p21 Waf1/Cip1, ultimately resulting in p53-mediated apoptosis rather than cell cycle arrest (4). Miz-1 also plays a role during lymphocyte development. In developing B and T cells, Miz-1 represses suppressor of cytokine signaling 1 (SOCS1) expression, which enables signaling through the IL-7 receptor and upregulation of the pro-survival protein Bcl-2 (5,6).

  1. Peukert, K. et al. (1997) EMBO J 16, 5672-86.
  2. Seoane, J. et al. (2001) Nat Cell Biol 3, 400-8.
  3. Staller, P. et al. (2001) Nat Cell Biol 3, 392-9.
  4. Seoane, J. et al. (2002) Nature 419, 729-34.
  5. Kosan, C. et al. (2010) Immunity 33, 917-28.
  6. Saba, I. et al. (2011) Blood 117, 3370-81.

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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