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.

    使用に関する制限

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    研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

    Revision 1
    #4487

    RNAi Machinery Antibody Sampler Kit

    RNAi Machinery Antibody Sampler Kit: Image 1 Expand Image
    Western blot analysis of extracts from various cell types using Argonaute 2 (C34C6) Rabbit mAb.
    RNAi Machinery Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from various cell types using Drosha (D28B1) Rabbit mAb.
    RNAi Machinery Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from various cell types using Argonaute 1 (D84G10) XP® Rabbit mAb.
    RNAi Machinery Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from MCF7 and 293 cells using Dicer (D38E7) Rabbit mAb.
    RNAi Machinery Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from mouse testis and mouse brain using Mili (D14F5) XP® Rabbit mAb (upper) or β-Actin Antibody #4967 (lower).
    RNAi Machinery Antibody Sampler Kit: Image 6 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    RNAi Machinery Antibody Sampler Kit: Image 7 Expand Image
    Immunoprecipitation of Argonaute 2 from COS-7 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Agonaute 2 (C34C6) Rabbit mAb. Western blot analysis was performed using Argonaute 2 (C34C6) Rabbit mAb.
    RNAi Machinery Antibody Sampler Kit: Image 8 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast using Argonaute 1 (D84G10) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
    RNAi Machinery Antibody Sampler Kit: Image 9 Expand Image
    Immunoprecipitation of Dicer from 293 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb XP Isotype Control #3900, and lane 3 is Dicer (D38E7) Rabbit mAb. Western blot analysis was performed using Dicer (D38E7) Rabbit mAb.
    RNAi Machinery Antibody Sampler Kit: Image 10 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse testis using Mili (D14F5) XP® Rabbit mAb.
    RNAi Machinery Antibody Sampler Kit: Image 11 Expand Image
    Confocal immunofluorescent analysis of mouse testis using Mili (D14F5) XP® Rabbit mAb (green) and Pan-Keratin (C11) Mouse mAb #4545 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).