Magna rip kit

Manufactured by Merck Group

Sourced in United States, Germany, China, Morocco, United Kingdom

The Magna RIP Kit is a laboratory equipment product designed for the extraction and purification of RNA-interacting proteins (RIPs) from biological samples. It provides a reliable and efficient method for isolating RIPs for further analysis and research purposes.

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Lab products found in correlation

Ab32381, by Abcam (19 mentions) Trizol reagent, by Thermo Fisher Scientific (16 mentions) Human anti ago2 antibody, by Merck Group (15 mentions) Ago2 antibody, by Abcam (13 mentions) Ago2 antibody, by Merck Group (13 mentions) Ab186733, by Abcam (12 mentions) Ab172730, by Abcam (11 mentions) Anti m6a antibody, by Synaptic Systems (10 mentions) Normal mouse igg, by Merck Group (10 mentions) Trizol, by Thermo Fisher Scientific (10 mentions) Anti ago2 antibody, by Abcam (9 mentions) Anti ago2, by Abcam (8 mentions) Anti ago2, by Merck Group (8 mentions) Rnase inhibitor, by Promega (8 mentions) Proteinase k, by Merck Group (8 mentions) Ago2 antibody, by Cell Signaling Technology (7 mentions) Magna rip rna binding protein immunoprecipitation kit, by Merck Group (7 mentions) Rip lysis buffer, by Merck Group (7 mentions) Anti m6a antibody, by Abcam (6 mentions) Trizol ls, by Thermo Fisher Scientific (6 mentions)

Spelling variants of this product

RIP kit (218 citations) Magna RIPTM RNA-Binding Protein Immunoprecipitation Kit (197 citations) EZ-Magna RIPTM RNA-Binding Protein Immunoprecipitation Kit (20 citations) Magna RIPTM RNA kit (6 citations) Magna Nuclear RIP kit (6 citations) Magna RIPTM RNA Immunoprecipitation Kit (4 citations) Z-Magna RIPTM RNA-binding Protein Immunoprecipitation kit (3 citations) EZ-Magna RIPTM Kit (3 citations) Magna RIPTM RNA binding protein co-immunoprecipitation kit (2 citations) Magna RIPTM RBP Immunoprecipitation Kit (2 citations)

692 protocols using magna rip kit

KIAA1429 Interactome and m6A Profiling

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RIP assay was conducted as previously described.³⁸ (link) Cell lysates were prepared with RIP lysis buffer (Magna RIP Kit; Millipore, USA) and then incubated with 5 μg of anti-KIAA1429 or rabbit IgG at 4°C overnight. The RNA-protein immunocomplexes were collected by protein A/G magnetic beads. After elution and purification, the purified RNA was analyzed by RT-PCR and qRT-PCR.
The m6A RIP was performed as previously described with some modifications.²⁷ (link) Total RNAs were isolated from SUM-1315 stable KIAA1429/METTL3 knockdown and control cells and treated with DNase I (Sigma Aldrich, USA). RNAs were fragmented by RNA fragmentation reagents. Immunoprecipitations were performed using an anti-m6A antibody (1:1,000; Abcam, USA) previously bound to magnetic Dynabeads (Life Technologies, USA) in the RIP Immunoprecipitation Buffer (Magna RIP Kit; Millipore, MA, USA) and incubated with DNA-free fragmented RNAs. RNAs were extracted by miRNeasy Mini kit (QIAGEN, Germany) and subjected to qRT-PCR and normalized to input.

Zhang X., Dai X.Y., Qian J.Y., Xu F., Wang Z.W., Xia T., Zhou X.J., Li X.X., Shi L., Wei J.F, & Ding Q. (2021). SMC1A regulated by KIAA1429 in m6A-independent manner promotes EMT progress in breast cancer. Molecular Therapy. Nucleic Acids, 27, 133-146.

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Immunoprecipitation-based RNA Interactome Profiling

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RIP assay was performed to identify the molecular interaction as previously described [14 (link)]. In brief, cells were lysed in complete RIP lysis buffer (Magna RIP Kit, Millipore, MA), and the isolated RNAs were fragmented by sonication and immunoprecipitated with protein A/G magnetic beads conjugated with specific antibodies (anti-METTL3, no. ab195352, Abcam; IGF2BP2, no. 11601–1-AP, Proteintech; anti-Flag, no. 8146, Cell Signaling) or control IgG in RIP Immunoprecipitation buffer (Magna RIP Kit, Millipore, MA) for 2 h at 4 °C. Beads were washed and incubated with Proteinase K to remove proteins. RNAs was extracted and subjected to qRT-PCR using primers and normalizing to input.

Guan H., Tian K., Luo W, & Li M. (2023). m6A-modified circRNA MYO1C participates in the tumor immune surveillance of pancreatic ductal adenocarcinoma through m6A/PD-L1 manner. Cell Death & Disease, 14(2), 120.

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Lysate Preparation and Immunoprecipitation for RNA Analysis

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Two lysis protocols were used in muscle biopsies and cultured cells: (1) skeletal muscle biopsies were cut into small pieces in RIP lysis buffer (Magna RIP Kit, Merck-Millipore, Burlington, MA) and homogenized by TissueLyser (Qiagen, Hilden, Germany); (2) Cultured cells were lysed with the following buffer: 150 mM KCl, 25 mM TrisHCl pH 7.4, 5 mM EDTA, 0.5% NP-40, 5 mM DTT, 1 mM PMSF, protease and RNAse inhibitors. For both lysates, homogenates were quantified for protein content, snap frozen in liquid nitrogen and stored at −80°C until use. For both muscle tissue and cells, 50 μg of lysate were used for total RNA analysis (INPUT RNA) and 1 mg was immunoprecipitated with anti-Ago2 or control mouse IgG antibodies (Abs) (RIPAb+ Ago2, Merck-Millipore, Burlington, MA), using 5 μg of antibody for 1 mg of lysate. Immunoprecipitation was performed using the Magna RIP Kit (Merck-Millipore, Burlington, MA) following the manufacturer’s instructions. IgG-bound RNA (IP RNA) was extracted from magnetic beads using TRIzol reagent (Invitrogen, ThermoFisher Scientific, Waltham, MA).

Cappella M., Perfetti A., Cardinali B., Garcia-Manteiga J.M., Carrara M., Provenzano C., Fuschi P., Cardani R., Renna L.V., Meola G., Falcone G, & Martelli F. (2018). High-throughput analysis of the RNA-induced silencing complex in myotonic dystrophy type 1 patients identifies the dysregulation of miR-29c and its target ASB2. Cell Death & Disease, 9(7), 729.

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QKI Isoform Immunoprecipitation Assay

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RNA immunoprecipitation assay was performed using the Magna Rip Kit (Millipore Sigma, # 17–700) per the manufacturer’s instructions using the following antibodies for QKI isoform pulldown: QKI-5 (rabbit anti-QKI, Millipore, # AB9904), QKI-6 (rabbit anti-QKI, Millipore, # AB9906), and QKI-7 (rabbit anti-QKI, Millipore, #AB9908).

Azam S.H., Porrello A., Harrison E.B., Leslie P.L., Liu X., Waugh T.A., Belanger A., Mangala L.S., Lopez-Berestein G., Wilson H.L., McCann J.V., Kim W.Y., Sood A.K., Liu J., Dudley A.C, & Pecot C.V. (2019). Quaking Orchestrates a Post-Transcriptional Regulatory Network of Endothelial Cell Cycle Progression Critical to Angiogenesis and Metastasis. Oncogene, 38(26), 5191-5210.

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Magnetic Bead Immunoprecipitation for RNA Analysis

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The Magna RIP™ Kit (MilliporeSigma®) was used following standard protocol. 10 µg of Antibody for Rabbit IgG (MilliporeSigma®, Cat.no.: PP64B) and hnRNPA2/B1 (Proteintech®, Cat.no.: 14813-1-AP) were used to load magnetic beads. RNA precipitate was subjected to qRT–qPCR analysis.

Feichtenschlager V., Chen L., Zheng Y.J., Ho W., Sanlorenzo M., Vujic I., Fewings E., Lee A., Chen C., Callanan C., Lin K., Qu T., Hohlova D., Vujic M., Hwang Y., Lai K., Chen S., Nguyen T., Muñoz D.P., Kohwi Y., Posch C., Daud A., Rappersberger K., Kohwi-Shigematsu T., Coppé J.P, & Ortiz-Urda S. (2024). The therapeutically actionable long non-coding RNA ‘T-RECS’ is essential to cancer cells’ survival in NRAS/MAPK-driven melanoma. Molecular Cancer, 23, 40.

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YTHDF2 RNA Immunoprecipitation Protocol

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RIP experiments were performed using the Magna RIP kit (MilliporeSigma) according to the instructions provided by the manufacturer. Briefly, the cells (1×10⁷ cells per 150 µl) were collected and lysed in a complete radioimmunoprecipitation assay buffer containing a protease inhibitor cocktail and RNase inhibitor. Magnetic beads coated with 5 µg specific antibodies against mouse IgG (SAB5600281, MilliporeSigma, Merck Co., Ltd.) or YTHDF2 (1/30, ab220163, Abcam) were pre-bound to protein A/G magnetic beads in immunoprecipitation buffer (20 mM Tris-HCl pH 7.5, 140 mM NaCl, 0.05% Triton X-100) (MilliporeSigma, Merck Co., Ltd.) for 2 h and then incubated with 100 µl cell lysate overnight at 4°C with rotation. RNA was eluted from the beads by incubation with 400 µl elution buffer for 2 h, precipitated with ethanol and dissolved in RNase-free water. The enrichment of certain fragments was determined using RT-qPCR.

Chen J.X., Chen D.M., Wang D., Xiao Y., Zhu S, & Xu X.L. (2023). METTL3/YTHDF2 m6A axis promotes the malignant progression of bladder cancer by epigenetically suppressing RRAS. Oncology Reports, 49(5), 94.

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Magna RIP™ Kit for RNA Binding

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The Magna RIP^™ Kit (MilliporeSigma^®) was used following standard protocol. 10μg of Antibody for Rabbit IgG (MilliporeSigma^®, Cat.no.: PP64B) and hnRNPA2/B1 (Proteintech^®, Cat.no.: 14813–1-AP) were used to load magnetic beads. RNA precipitate was subjected to qRT–qPCR analysis.

Feichtenschlager V., Chen L., Zheng Y.J., Ho W., Sanlorenzo M., Vujic I., Fewings E., Lee A., Chen C., Callanan C., Lin K., Qu T., Hohlova D., Vujic M., Hwang Y., Lai K., Chen S., Nguyen T., Muñoz D.P., Kohwi Y., Posch C., Daud A., Rappersberger K., Kohwi-Shigematsu T., Coppé J.P, & Ortiz-Urda S. (2023). The therapeutically actionable long non-coding RNA ‘T-RECS’ is essential to cancer cells’ survival in NRAS/MAPK-driven melanoma. Research Square.

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XIST-miR-34a Interaction Analysis via RIP Assay

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The interaction between XIST and miR-34a was investigated by RIP assays using a Magna RIP kit (MilliporeSigma; Merck KGaA, Darmstadt, Germany), according to the manufacturer's instructions. Briefly [28 (link)], RIPA lysis buffer was used to lyse AML cells (∼2 × 10⁷ cells). The supernatant was collected for RIP assays using an EZ-Magna RIPTM RNA-Binding Protein Immunoprecipitation Kit (MilliporeSigma). 10% cell extract was used as the input, and 100 μL cell extract was incubated with 50 μL protein A/G conjugated magnetic beads precoated with antibodies against Argonaute2 (Ago2) or IgG at 4 °C overnight. TRIzol was used for RNA extraction, and RT-qPCR was used for detection. The expression of XIST and miR-34a was analyzed in the anti-Ago2 and anti-IgG groups, respectively.

Wen C., Lu X., Sun Y., Li Q., Liao J, & Li L. (2023). Naringenin induces the cell apoptosis of acute myeloid leukemia cells by regulating the lncRNA XIST/miR-34a/HDAC1 signaling. Heliyon, 9(5), e15826.

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Magna RIP™ Kit Antibody Pulldown

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The Magna RIP™ Kit (MilliporeSigma®) was used following standard protocol. 10µg of Antibody for Rabbit IgG (MilliporeSigma®, Cat.no.: PP64B) and hnRNPA2/B1 (Proteintech®, Cat.no.: 14813-1-AP) were used to load magnetic beads. RNA precipitate was subjected to qRT-qPCR analysis.

Feichtenschlager V., Chen L., Zheng Y.J., Ho W., Sanlorenzo M., Vujic I., Fewings E., Lee A., Chen C.S., Lin K., Callanan C., Vujic M., Hwang Y., Lai K., Chen S., Nguyen T., Muñoz D.P., Kohwi Y., Posch C., Daud A., Rappersberger K., Kohwi-shigematsu T., Coppé J., & Ortiz‐Urda S. (2022). The long non-coding RNA ‘TRASH’ is essential for cell survival in MAPK-driven melanoma.

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RIP Assay for EZH2 Interactome

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Magna RIP kit and DZNep were purchased from Millipore Sigma (Burlington, MA). RIP was performed according to manufacturer’s instructions. Briefly, cell lysates were incubated with antibody-bead mix overnight at 4°C. Following this, proteins were degraded and the RNA was isolated by the phenol-chloroform extraction method. The RNA was precipitated using Kit components and ethanol at −80°C overnight. For EZH2 inhibition, cells were treated with 500 μM DZNep for 72 h before harvesting for RIP assay.

Vaidya A.M., Sun Z., Ayat N., Schilb A., Liu X., Jiang H., Sun D., Scheidt J., Qian V., He S., Gilmore H., Schiemann W.P, & Lu Z.R. (2019). Systemic Delivery of Tumor-Targeting siRNA Nanoparticles against an Oncogenic LncRNA Facilitates Effective Triple-Negative Breast Cancer Therapy. Bioconjugate chemistry, 30(3), 907-919.

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