Typically, one RIP reaction (i.e., one immunoprecipitation using one antibody) requires 100 ?L of cell lysate from approximately 2.0 x 10^7 cells or one 15 cm plate. However, the total number of cells or total amount of protein used per RIP must be optimized based on the abundance of the RNA-binding protein being investigated and the planned method of RNA detection.
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Magna RIP® RNA-Binding Protein Immunoprecipitation Kit
RNA Immunoprecipitation (RIP) Kit containing all necessary reagents to perform 12 individual RNA-binding protein immunoprecipitation (RIP) reactions using protein A/G magnetic beads.
Magna RIP® RNA-Binding Protein Immunoprecipitation Kit
Synonym(s):
Magnetic Bead RNA Immunoprecipitation, Magnetic RIP kit, RIP Kit, RNA Immunopreciptation
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About This Item
UNSPSC Code:
12161503
NACRES:
NA.32
eCl@ss:
32161000
Pricing and availability is not currently available.
Quality Segment
manufacturer/tradename
Magna RIP®, Upstate®
technique(s)
RIP: suitable, immunoprecipitation (IP): suitable
shipped in
dry ice
General description
Gene regulation plays a critical role in complex cellular processes such as development, differentiation, and cellular response to environmental changes. In addition to transcriptional regulation of gene expression by transcription factors, cells utilize post-transcriptional regulatory mechanisms. One such mechanism involves use of certain RNA-binding proteins (RBPs) to temporally and coordinately regulate the rate of mRNA translation of functionally related gene
products. While the regulation of gene expression by transcription factors has been well studied over time, the post-transcriptional regulation of mRNAs by RBPs and the role of non-coding RNAs in this process is a relatively nascent field that remains to be thoroughly explored.
products. While the regulation of gene expression by transcription factors has been well studied over time, the post-transcriptional regulation of mRNAs by RBPs and the role of non-coding RNAs in this process is a relatively nascent field that remains to be thoroughly explored.
RNA-binding protein immunoprecipitation (RIP) is the RNA analog of the more well-known ChIP application (chromatin immunoprecipitation), which identifies DNA targets of DNA-binding proteins in an in-vivo cellular context. RIP can be used to identify specific RNA molecules (of many types) associated with specific nuclear or cytoplasmic binding proteins. These experiments involve immunoprecipitation of endogenously formed complexes of RNA-binding proteins and co-isolation of any RNA species associated with that RNA-binding protein. Purification of these RNA species allows interrogation and identification of mRNAs (and potentially non-coding RNAs associated with them) and can be directly measured using down stream applications including quantitative reverse transcription polymerase chain reaction (RT-PCR), microarray analysis (RIP-chip) and “deep-sequencing” or 2nd-generation sequencing based platforms (RIP-Seq).
Features & Benefits:
-Protein A/G magnetic beads, optimized to bind nucleic acid-protein immune complexes
-RNAse inhibitors and RNAse-free reagents
-Negative controls
Features & Benefits:
-Protein A/G magnetic beads, optimized to bind nucleic acid-protein immune complexes
-RNAse inhibitors and RNAse-free reagents
-Negative controls
Application
Research Category
Epigenetics & Nuclear Function
Epigenetics & Nuclear Function
Packaging
RIP Kit capacity: 12 RNA-binding protein immunoprecipitation assays
Physical form
Two boxes containing all necessary reagents to perform 12 individual RNA-binding protein immunoprecipitation (RIP) reactions.
Preparation Note
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.
Other Notes
Magnetic Beads Protein A/G
RIP Wash Buffer
RIP Lysis Buffer
0.5 M EDTA
10% SDS
Salt Solution I
Salt Solution II
Precipitate Enhancer
Normal Mouse IgG
Rabbit IgG Purified
Protease Inhibitor Cocktail 200X
RNase Inhibitor
Proteinase K (10 mg/mL)
Nuclease free water
RIP Wash Buffer
RIP Lysis Buffer
0.5 M EDTA
10% SDS
Salt Solution I
Salt Solution II
Precipitate Enhancer
Normal Mouse IgG
Rabbit IgG Purified
Protease Inhibitor Cocktail 200X
RNase Inhibitor
Proteinase K (10 mg/mL)
Nuclease free water
Legal Information
MAGNA RIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
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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This Item | |||
|---|---|---|---|
| technique(s) RIP: suitable, immunoprecipitation (IP): suitable | technique(s) RIP: suitable, activity assay: suitable (protein interaction), immunoprecipitation (IP): suitable | technique(s) - | technique(s) immunoprecipitation (IP): suitable |
| manufacturer/tradename Magna RIP®, Upstate® | manufacturer/tradename Magna Nuclear RIP | manufacturer/tradename - | manufacturer/tradename Magna ChIP® |
| Quality Level 100 | Quality Level 100 | Quality Level 200 | Quality Level 100 |
| shipped in dry ice | shipped in dry ice | shipped in - | shipped in dry ice |
Storage Class
10 - Combustible liquids
target_organs
Respiratory Tract
wgk
WGK 3
signalword
Danger
Hazard Classifications
Acute Tox. 4 Oral - Aquatic Chronic 3 - ED ENV 1 - Eye Irrit. 2 - Skin Irrit. 2 - STOT RE 2 Inhalation
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Could you advise on the maximum number of cells to start with to ensure high yield and purity?
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Is an antibody suitable for Co-IP also suitable for RIP? Is there any difference between hosts/immunoglobulin affinity?
1 answer-
An antibody suitable for co-immunoprecipitation (Co-IP) of proteins is not necessarily suitable for RNA immunoprecipitation (RIP). Regardless of host, an antibody suitable for RIP targets accessible epitopes on the RNA binding protein of interest, which are not masked by the interaction with target RNAs. The amount of antibody used for RIP depends on the presentation (e.g., purified or unpurified) and the effective affinity of the candidate antibody when used for immunoprecipitation. It is recommended to use RIP-validated antibodies, such as RIPAb+ validated antibodies. Examples can be found at the following link: https://www.sigmaaldrich.com/search/ripab%2B-validated-antibody?focus=products&page=1&perpage=30&sort=relevance&term=RIPAb%2B%20validated%20antibody&type=product
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For RIP-seq, should I perform rRNA depletion after RNA purification？
1 answer-
Assuming rRNA are not known targets and the target RNA has been enriched using the binding protein/antibody, a depletion step is not required.
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Hi, on the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit, after RNA insolation do RNA samples need DNAseI treatment to eliminate DNA contaminants? Or DNA was eliminated in a different way? Thanks Martin
1 answer-
The need for DNA removal from RIPed RNA can vary depending on the specific RNA binding protein targets. Typically, most RIPed RNA samples do not require DNA removal. However, if DNA contamination of the sample is anticipated, it is recommended to perform DNase I treatment or include a minus RT (reverse transcription) sample for the qPCR analysis.
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