Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

SignalSilence® p27 Kip1 siRNA I #12324

Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
TFN Human,

Species cross-reactivity is determined by western blot.

Applications Key: TFN=Transfection,


Species predicted to react based on 100% sequence homology: Monkey,

Specificity / Sensitivity

SignalSilence® p27 Kip1 siRNA I inhibits human and monkey p27 Kip1 expression. SignalSilence® p27 Kip1 siRNA I可以抑制人和猴的p27 Kip1蛋白表达。


SignalSilence® p27 Kip1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit p27 Kip1 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.Cell Signaling Technology (CST) 公司的SignalSilence® p27 Kip1 siRNA I 帮助研究人员使用RNA干扰技术特异性抑制p27 Kip1表达,该技术通过向细胞内导入双链RNA分子以选择性的抑制某些基因的表达。所有CST公司的SignalSilence® siRNA都经过了严格的内部测试,western分析显示能够能够降低目标蛋白的表达。

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. 寡核苷酸合成经过了三苯甲基的逐碱基分析以确保耦合效率。随后经过亲和固相萃取纯化。退火的RNA双链在经过质谱分析以确保双链的准确性。新批次都会经过质谱分析与之前的批次进行比对以保证不同批次之间的稳定性。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® p27 Kip1 siRNA I (+), or SignalSilence® p27 Kip1 siRNA II #12410 (+), using p27 Kip1 (D69C12) XP® Rabbit mAb #3686 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The p27 Kip1 (D69C12) XP® Rabbit mAb confirms silencing of p27 Kip1 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.转染100 nM SignalSilence®对照siRNA(未标记) #6568 (-),SignalSilence® p27 Kip1 siRNA I (+),或SignalSilence® p27 Kip1 siRNA II #12410 (+)的HeLa细胞提取物,使用p27 Kip1 (D69C12) XP® Rabbit mAb #3686 (上)或β-Actin (D6A8) Rabbit mAb #8457 (下)进行western blot分析。p27 Kip1 (D69C12) XP® Rabbit mAb可以确认p27 Kip1表达受抑制,β-Actin (D6A8) Rabbit mAb用于确认上样量一致。

Directions for Use

CST recommends transfection with 100 nM SignalSilence® p27 Kip1 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. CST建议转染100 nM SignalSilence® p27 Kip1 siRNA I,48-72小时后裂解细胞。转染的步骤请参阅转染试剂的说明书。关于使用问题,可以随时咨询CST。 每管试剂能进行100次转染,该转染次数以总体积为300μl 的1个24孔板孔中siRNA终浓度100 nM来进行计算。


p27 Kip1 is a member of the Cip/Kip family of cyclin-dependent kinase inhibitors. Like its relatives, p57 Kip2 and p21 Waf1/Cip1, the ability to enforce the G1 restriction point is derived from its inhibitory binding to CDK2/cyclin E and other CDK/cyclin complexes. Expression levels of p27 are upregulated in quiescent cells and in cells treated with cAMP or other negative cell cycle regulators. Downregulation of p27 can be induced by treatment with interleukin-2 or other mitogens; this involves phosphorylation of p27 and its degradation by the ubiquitin-proteasome pathway (1-4).p27 Kip1是细胞周期蛋白激酶抑制因子Cip/Kip家族的一员。与它的同类蛋白类似,p57 Kip2 和p21 Waf1/Cip1,它实施G1限制点的能力来自于阻止和CDK2/cyclin E和其他CDK/cyclin复合物结合。p27的表达水平在休眠细胞和被cAMP或其他细胞周期负调因子处理过的细胞中表达上调。白介素2或其他有丝分裂原处理后能够下调p27;这一机制设计了p27的磷酸化,p27的降解是通过泛素-蛋白酶体通路进行的(1-4)。

  1. Lloyd, R.V. et al. (1999) Am J Pathol 154, 313-23.
  2. Polyak, K. et al. (1994) Genes Dev 8, 9-22.
  3. Kato, J.Y. et al. (1994) Cell 79, 487-96.
  4. Vlach, J. et al. (1997) EMBO J 16, 5334-44.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

SignalSilence is a registered trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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