The RIP assay was carried out with the RIP™ RNA-Binding Protein Immunoprecipitation Kit from EMD Millipore. Briefly, SiHa cells transfected with miR-129-5p NC or inhibitor were collected. Then, cell lysate was incubated with RIP buffer containing magnetic beads conjugated with an anti-Ago2 antibody or IgG isotype control. Afterwards, RT-qPCR analysis was employed to evaluate the immunoprecipitated RNA.
Rip rna binding protein immunoprecipitation kit
Manufactured by Merck Group
Sourced in United States, United Kingdom
The RIP RNA-Binding Protein Immunoprecipitation Kit is a tool for the study of RNA-protein interactions. It is used to isolate and identify RNA molecules that are associated with specific RNA-binding proteins within a cell.
Automatically generated - may contain errors
Lab products found in correlation
Anti ago2 antibody, by Abcam (3 mentions)
Rabbit anti hnrnpa1 antibody, by Cell Signaling Technology (2 mentions)
Streptavidin c1 magnetic beads, by Thermo Fisher Scientific (1 mentions)
Pierce magnetic rna protein pull down kit, by Thermo Fisher Scientific (1 mentions)
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
Gfp antibody ab290, by Abcam (1 mentions)
Anti ago2 or anti igg antibody, by Abcam (1 mentions)
Chip assay kit, by Merck Group (1 mentions)
Dnmt3a, by Abcam (1 mentions)
Ago2 antibody, by Merck Group (1 mentions)
Complete rip lysis buffer, by Merck Group (1 mentions)
Normal mouse igg, by Merck Group (1 mentions)
Human anti argonaute2 anti ago2 antibody, by Merck Group (1 mentions)
Ago2 conjugated magnetic beads, by Merck Group (1 mentions)
Anti argonaute 2 antibody ago2, by Abcam (1 mentions)
M 280 streptavidin magnetic bead, by Thermo Fisher Scientific (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Rneasy minelute cleanup kit, by Qiagen (1 mentions)
Anti ago2 antibody, by Merck Group (1 mentions)
Primescript rt master mix, by Takara Bio (1 mentions)
43 protocols using rip rna binding protein immunoprecipitation kit
1
Ago2-RNA Immunoprecipitation Assay
2
Biotinylated LINC00341 Interactome Identification
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The biotinylated LINC00341, antisense LINC00341, or mutated LINC00341 was mixed with proteins obtained from cancer cells for overnight at 4°C. The complex of biotinylated lncRNA and proteins was purified using streptavidin‐agarose for 4 hours at 4°C. The proteins are then eluted from the RNA‐protein complex and detected by Western blotting analysis. RIP assay was performed using RIP RNA‐binding protein immunoprecipitation kit (Merck Millipore) and following the manufacturer's protocol.
3
RNA-Binding Protein HuR Enrichment
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For RNA pull‐down assay, the biotin‐labelled probes were designed and synthesized (GenePharma) and then incubated with the lysates of 2 × 107 treated Hep‐2 and TU212 cells overnight at 37°C, followed by addition of C1 streptavidin magnetic beads (Invitrogen) for 2 hours at 25°C. The protein enriched by probe was subjected to Western blot analysis for HuR expression. RIP assay was conducted with 5 μg anti‐HuR antibody (#12582, CST) using the RIP™ RNA‐Binding Protein Immunoprecipitation Kit (Merck Millipore) in accordance with the manufacturer's instructions.
4
RIP-qRT-PCR Analysis of miR-195-5p
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The experiment was implemented with the assistance of the RIP™ RNA-Binding Protein Immunoprecipitation Kit (Merck Millipore). RIP lysis buffer was administered to lyse LoVo and LS513 cells, and RNA (miR-NC or miR-195-5p) magnetic beads were combined with the mouse anti-IgG antibody and the human anti-Ago2 antibody or human anti-IGF2BP2 antibody [34 (link)]. TUG1’s level was determined through qRT-PCR.
5
RNA-Protein Interaction Analysis of CircMMP9
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RNA pull-down assay was performed using PierceTM Magnetic RNA-Protein
Pull-Down Kit (#20164, Thermo Fisher Scientific, Waltham, MA, USA) with biotinylated
control or circ-MMP9 probe in UM1 and HSC-3 cells, followed by Western blot analysis for
AUF1 protein expression and quantitative real-time PCR (qRT-PCR) analysis for miR-183 and
miR-149 expression. Radioimmunoprecipitation assay (RIPA) was carried out using RIP™
RNA-Binding Protein Immunoprecipitation Kit (Merck Millipore, Darmstadt, Germany) in
accordance with the manufacturer’s protocols, followed by qRT-PCR analysis for circ-MMP9
expression.
Pull-Down Kit (#20164, Thermo Fisher Scientific, Waltham, MA, USA) with biotinylated
control or circ-MMP9 probe in UM1 and HSC-3 cells, followed by Western blot analysis for
AUF1 protein expression and quantitative real-time PCR (qRT-PCR) analysis for miR-183 and
miR-149 expression. Radioimmunoprecipitation assay (RIPA) was carried out using RIP™
RNA-Binding Protein Immunoprecipitation Kit (Merck Millipore, Darmstadt, Germany) in
accordance with the manufacturer’s protocols, followed by qRT-PCR analysis for circ-MMP9
expression.
6
RIP Assay for RNA-Protein Interactions
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The RIP assay was performed according to the instructions of the RIP RNA-binding protein Immunoprecipitation kit (EMD Millipore) following the manufacturer's protocol. The PC cell lysates were added with RIP Lysis Buffer and NELFE antibody, normal mouse IgG (negative control) (1:200; cat. no. PP6421-K; EMD Millipore) and agarose beads and incubated overnight at 4°C. A High-Capacity cDNA Reverse Transcription kit (Thermo Fisher Scientific, Inc.) was used to reverse transcribe the immunoprecipitated RNAs. Finally, RT-qPCR was performed as aforementioned in order to examine the targets transcripts.
7
Detecting SNHG22 RNA-Protein Interactions
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RIP™ RNA-Binding Protein Immunoprecipitation Kit (Millipore, USA) was used for detecting the binding between SNHG22 and EZH2 or SNHG22 and Ago2. Briefly, GC cell lysates were incubated with magnetic beads conjugated with indicated antibody or IgG as a negative control. Followed qRT-PCR analysis was performed to detect the enrichment of SNHG22.
8
AGO2-Bound RNA Identification
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We performed the RIP assays with a RIP™ RNA-binding protein immunoprecipitation kit (Millipore, Bedford, MA, USA) according to the manufacturer’s instructions. A total of 2 × 107 cells were lysed with the RIP lysis buffer. Then, anti-AGO2 antibody or negative control anti-IgG antibody was incubated with magnetic beads for 30 min. Subsequently, we mixed the magnetic beads with cell lysate and incubated overnight at 4°C. Finally, we isolated the AGO2-binding RNA for the following qRT-PCR.
9
Argonaute2 Immunoprecipitation in Glioma
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LN229 and N3 glioma cells were treated with magnetic beads pre-conjugated with Argonaute2 (Ago2) antibody. Normal mouse IgG functioned as a negative control. RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore, Billerica, MA, USA) was utilized for RIP assay in accordance with the manufacturer’s protocol.
10
Ago2 Immunoprecipitation and RIP
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Briefly, magnetic beads were pre-conjugated with antibody against Argonaute2 (Ago2). Related LN229 and U251 glioma cell lysates were harvested and treated with bead-antibody complex. Normal mouse IgG was used as a negative control. RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore, Billerica, MA, USA) was chosen for RIP and all experiments were performed according to manufacturer’s instructions.
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