Revision 1

#4487Store at -20C

1 Kit

(5 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Drosha (D28B1) Rabbit mAb 3364 20 µl 160 kDa Rabbit IgG
Argonaute 2 (C34C6) Rabbit mAb 2897 20 µl 97 kDa Rabbit IgG
Argonaute 1 (D84G10) XP® Rabbit mAb 5053 20 µl 97 kDa Rabbit IgG
Mili (D14F5) XP® Rabbit mAb 5940 20 µl 110 kDa Rabbit IgG
Dicer (D38E7) Rabbit mAb 5362 20 µl 220 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The RNAi Machinery Antibody Sampler Kit provides an economical means to analyze proteins associated with endogenous RNA interference. The kit contains enough primary and secondary antibodies to perform two western blot experiments.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

RNA interference (RNAi) serves as a global mechanism of gene regulation in eukaryotes. Through interactions with Dicer, Drosha, Argonaute 2 (Ago2) and Miwi/Mili proteins, microRNA (miRNA) is processed within the nucleus and utilized for gene silencing and down regulation of gene expression. Dicer is a member of the RNase III family that specifically cleaves double-stranded RNA to generate microRNA (miRNA) (1). Long, primary transcripts (pri-miRNAs) are processed to stem-looped pre-miRNAs by the nuclear RNase III Drosha (2) and are then transported to the cytoplasm for further processing by Dicer to produce mature, 22-nucleotide miRNAs (3). The mature miRNA then becomes a part of the RNA-Induced Silencing Complex (RISC) and can bind to the 3' UTR of the target mRNA (3). Interference of Drosha pri-miRNA processing results in the increase of pri-miRNAs and the decrease of pre-miRNAs (2). Drosha forms part of a multiprotein complex called the Microprocessor along with other components, such as DGCR8 (4). Both Drosha and DGCR8 are necessary for miRNA biogenesis (5). Argonaute protein family members participate in various steps of miRNA-mediated gene silencing such as repression of translation and mRNA turnover (6). The Drosophila piwi gene was identified as being required for the self-renewal of germ-line stem cells, and its homologues are well conserved among various species including Arabidopsis, C. elegans and human (7). Miwi and Mili proteins are both mouse homologs of Piwi and contain a carboxy-terminal Piwi domain that binds to Piwi-interacting RNAs (piRNAs) in male germ cells and are essential for spermatogenesis in mouse (8-11).

  1. Hutvágner, G. and Zamore, P.D. (2002) Science 297, 2056-60.
  2. Lee, Y. et al. (2003) Nature 425, 415-9.
  3. Diederichs, S. and Haber, D.A. (2007) Cell 131, 1097-108.
  4. Denli, A.M. et al. (2004) Nature 432, 231-5.
  5. Gregory, R.I. et al. (2004) Nature 432, 235-40.
  6. Peters, L. and Meister, G. (2007) Mol Cell 26, 611-23.
  7. Cox, D.N. et al. (1998) Genes Dev 12, 3715-27.
  8. Kuramochi-Miyagawa, S. et al. (2001) Mech Dev 108, 121-33.
  9. Aravin, A. et al. (2006) Nature 442, 203-7.
  10. Grivna, S.T. et al. (2006) Proc Natl Acad Sci U S A 103, 13415-20.
  11. Grivna, S.T. et al. (2006) Genes Dev 20, 1709-14.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

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