Thermo Scientific™ Pierce™ Magnetic RNA-Protein Pull-Down Kit

Thermo Scientific™ Pierce™ Magnetic RNA-Protein Pull-Down Kit

The Thermo Scientific Pierce Magnetic RNA-Protein Pull-Down Kit provides researchers with a streamlined, robust method to enrich protein-RNA interactions using end-labeled RNA as the bait.

The Magnetic RNA-Protein Pull-Down Kit provides reagents to efficiently enrich RNA Binding Proteins (RBPs) using RNA end-labeled with desthiobiotin and streptavidin magnetic beads. The complete kit contains sufficient reagents for 20 RNA labeling reactions and 20 protein-RNA pull-down assays. This direct enrichment of the protein-RNA interaction provides an alternative to antibody capture of protein-RNA complexes or moieties incorporated into the nucleic acid. An added advantage to the kit is that it includes validated controls for both the labeling and pull-down assay. The kit is amenable to several downstream applications, including Western blotting and Mass Spectrometry (MS).

Features

• Direct – capture ribonucleoprotein complexes directly with end-labeled RNA; does not use or require antibodies for pull-down
• Easy to use – no spin cups or centrifugation necessary for the enrichment of RBP; procedure streamlined for minimal hands-on time (less than 3 hours) after RNA labeling reaction
• Flexible – use in vitro transcribed RNA or synthetic RNA for labeling of various lengths and complexity; proteins successfully enriched using endogenous, over-expressed, and in vitro translated lysates
• Specific – low bead background; unrelated RNA or mutated RNA does not significantly enrich specified RBPs
• Economical – less expensive than purchasing commercially synthesized end-labeled RNA, magnetic beads, and reagents separately
• Complete – contains both labeling and enrichment modules with buffers necessary for assay; positive control RNA, negative control RNA, and RBP antibody included

Contents

Sufficient For: 20 RNA-protein complex pull-down reactions

• Pierce Nucleic Acid-Compatible Streptavidin Magnetic Beads, 1 mL (store at 4°C)
• RNA Capture Buffer (1X), 10 mL (store at 4°C)
• Tris (20 mM, pH 7.5), 5 mL (store at 4°C)
• Protein-RNA Binding Buffer (10X), 1 mL (store at 4°C)
• Wash Buffer (1X), 10 mL (store at 4°C)
• Biotin Elution Buffer, 1.5 mL (store at 4°C)
• HuR Monoclonal Antibody (Mouse), 50 µL (store at 4°C)
• Positive RNA Control (AR RNA), 250 pmol (store at -20°C)
• Negative RNA Control (polyA25 RNA), 250 pmol (store at -20°C)
• Pierce RNA 3' End Desthiobiotinylation Kit (part number 20163; store at -20°C)