Typically, CLIP assays require that samples be UV cross-linked. As such, the RIP kit 17-700 is not designed for samples that have been UV cross-linked and would not be suitable for CLIP assays.
Wybierz wielkość
| Rozmiar/SKU | Dostępność | Cena netto |
|---|---|---|
12 assays | Skontaktuj się z Obsługą Klienta, aby uzyskać informacje na temat dostępności | 3480,00 zł |
Informacje o tej pozycji
3480,00 zł
Quality Segment
manufacturer/tradename
Magna RIP®, Upstate®
technique(s)
RIP: suitable, immunoprecipitation (IP): suitable
shipped in
dry ice
General description
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.
Features & Benefits:
-Protein A/G magnetic beads, optimized to bind nucleic acid-protein immune complexes
-RNAse inhibitors and RNAse-free reagents
-Negative controls
Application
Epigenetics & Nuclear Function
Packaging
Physical form
Preparation Note
Other Notes
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
Disclaimer
Klasa składowania
10 - Combustible liquids
Narządy docelowe
Respiratory Tract
Co się dzieje?
WGK 3
Słowo ostrzegawcze
Danger
Kody zagrożeń
Ostrzeżenia
Hazard Classifications
Acute Tox. 4 Oral - Aquatic Chronic 3 - ED ENV 1 - Eye Irrit. 2 - Skin Irrit. 2 - STOT RE 2 Inhalation
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Can I use this kit to perform CLIP assay?
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Could you advise on the maximum number of cells to start with to ensure high yield and purity?
1 answer-
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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Is an antibody suitable for Co-IP also suitable for RIP? Is there any difference between hosts/immunoglobulin affinity?
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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?
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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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