Revision 1

#4487

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RNAi Machinery Antibody Sampler Kit

1 Kit

(5 x 20 microliters)

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
Drosha (D28B1) Rabbit mAb336420 µl160 kDaRabbit IgG
Argonaute 2 (C34C6) Rabbit mAb289720 µl97 kDaRabbit IgG
Argonaute 1 (D84G10) XP® Rabbit mAb505320 µl97 kDaRabbit IgG
Mili (D14F5) XP® Rabbit mAb594020 µl110 kDaRabbit IgG
Dicer (D38E7) Rabbit mAb536220 µl220 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

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).

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

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

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Western blot analysis of extracts from various cell types using Argonaute 2 (C34C6) Rabbit mAb.

undefined Image 1: RNAi Machinery Antibody Sampler Kit

Western blot analysis of extracts from various cell types using Drosha (D28B1) Rabbit mAb.

undefined Image 2: RNAi Machinery Antibody Sampler Kit

Western blot analysis of extracts from various cell types using Argonaute 1 (D84G10) XP® Rabbit mAb.

undefined Image 3: RNAi Machinery Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

CST Logo

Western blot analysis of extracts from MCF7 and 293 cells using Dicer (D38E7) Rabbit mAb.

undefined Image 4: RNAi Machinery Antibody Sampler Kit

Western blot analysis of extracts from mouse testis and mouse brain using Mili (D14F5) XP® Rabbit mAb (upper) or β-Actin Antibody #4967 (lower).

undefined Image 5: RNAi Machinery Antibody Sampler Kit

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.

undefined Image 6: RNAi Machinery Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

CST Logo

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.

undefined Image 7: RNAi Machinery Antibody Sampler Kit

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).

undefined Image 8: RNAi Machinery Antibody Sampler Kit

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.

undefined Image 9: RNAi Machinery Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

CST Logo

Immunohistochemical analysis of paraffin-embedded mouse testis using Mili (D14F5) XP® Rabbit mAb.

undefined Image 10: RNAi Machinery Antibody Sampler Kit

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).

undefined Image 11: RNAi Machinery Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.