EZ-Magna NuCLEAR^™ RIP (Cross-Linked) Nuclear RNA-Binding Protein Immunoprecipitation Kit

Features and Advantages

• Generates cross-linked chromatin to allow analysis of a variety of chromatin-associated RNAs
• Flexible, scalable input requirements: Recover RNA from millions of cells or as few as 5,000 cells
• Magnetic protein A/G bead blend and optimized buffer system results in low backgrounds and high signal-to-noise ratios
• Suitable for analysis by RT-qPCR or RIP-seq
• Complete set of reagents and detailed protocol to enable first-time success

Magna Nuclear RIP Kits are specially designed to allow the discovery and analysis of a variety of chromatin associated RNAs such as long non-coding RNAs, enhancer RNAs and miRNAs . These chromatin-associated RNAs often regulate gene expression and can be analyzed with applications including quantitative reverse transcription polymerase chain reaction (RT-PCR), microarray analysis (RIP-chip) and next generation sequencing (RIP-Seq).

Nuclear RIP can be performed using chromatin that has interactions stabilized by formaldehyde treatment (cross-linked) or chromatin that has not been treated with a cross linking reagent (native). While both of these approaches are similar in that they are designed to recover chromatin associated RNA, the reagents used and the details of the protocol and types of interactions typically detected are different. Cross-linked can capture higher molecular weight complexes in in vivo configurations with possibly lower affinities. In contrast native RIP is expected to recover high affinity, more direct interactions between proteins encoded RNA binding motifs and candidate RNAs. For less well understood proteins and protein complexes often both approaches are used.

The kit described here is for the cross-linked approach. If a native approach is of interest please visit the product page for the Magna Nuclear RIP (Native) Kit, catalogue # 17-10522 or the EZ-Magna Nuclear RIP (Native) Kit, catalogue # 17-10523.

Kit capacity: 24 RNA-binding protein immunoprecipitation assays using a cross-linked nuclear lysate

Two boxes containing key reagents for generation of cross linked nuclear lysates and performance of 24 individual RNA-binding protein immunoprecipitation (RIP) reactions. Plus box containing positive and negative control antibodies and qPCR primers.

Upon receipt, store components at the temperatures indicated on the labels.
Kit components are stable for 6 months from date of shipment when stored as directed.

10X Glycine

10X PBS

Nuclei Isolation Buffer

RIP Cross-Linked Lysis Buffer

Protein A/G Magnetic Beads

Nuclear RIP Dilution Buffer

Low Salt Wash Buffer

High Salt Wash Buffer

LiCl Wash Buffer

TE Buffer

RIP Elution Buffer

10% SDS

0.5 M EDTA

DNase I (RNase Free)

DNase I Supplement

DNase I Reaction Buffer

Protease Inhibitor Cocktail III, Animal Free

RNAse Inhibitor

Proteinase K


Control Antibodies and Primers
Normal Mouse IgG Negative Control Antibody

Anti-EZH2 Positive Control Antibody

NEAT1 Positive Control Primers

U1 snRNA Negative Control Primers

NuCLEAR is a trademark of Sigma-Aldrich Co. LLC