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EZ-Magna NuCLEAR™ RIP (Native) Nuclear RNA-Binding Protein Immunoprecipitation Kit
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General description
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. Native RIP is expected to recover high affinity, more direct interactions between proteins encoded RNA binding motifs and candidate RNAs. In contrast, nuclear RIP with cross-linked chromatin can capture higher molecular weight complexes in in vivo configurations with possibly lower affinities. For less well understood proteins and protein complexes often both approaches are used.
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. Native RIP is expected to recover high affinity, more direct interactions between proteins encoded RNA binding motifs and candidate RNAs. In contrast, nuclear RIP with cross-linked chromatin can capture higher molecular weight complexes in in vivo configurations with possibly lower affinities. For less well understood proteins and protein complexes often both approaches are used.
Application
EZ-Magna Nuclear RIP (Native) RNA-Binding Protein Immunoprecipitation Kit is designed for the analysis of chromatin associated RNA such lncRNAs, enhancer RNAs and miRNAs.
Packaging
Kit capacity: 24 RNA-binding protein immunoprecipitation assays using a native (non-cross-linked) nuclear lysate
Components
Nuclei Isolation Buffer ;RIP Lysis Buffer;Protein A/G Magnetic Beads;RIP Lysis Buffer;0.5M EDTA;DNase I (RNase Free) 2 U/μL;DNase I Reaction Buffer ;Protease Inhibitor Cocktail III, Animal Free;RNAse Inhibitor;Control Antibodies and PrimersNormal Mouse IgG Negative Control Antibody;Anti-EZH2 Positive Control Antibody ;NEAT1 Positive Control Primers ;U1 snRNA Negative Control Primers
Legal Information
NuCLEAR is a trademark of Sigma-Aldrich Co. LLC
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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Hazard Classifications
Aquatic Chronic 3 - Eye Irrit. 2 - Skin Irrit. 2
Storage Class
10 - Combustible liquids
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WGK 3
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