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Imprint rip kit

Manufactured by Merck Group
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The Imprint® RIP kit is a laboratory equipment product manufactured by Merck Group. It is designed to facilitate the isolation and purification of recombinant proteins expressed in Escherichia coli or other bacterial host cells. The kit provides the necessary reagents and protocols to enable the effective recovery and concentration of target proteins from cell lysates.

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Lab products found in correlation

High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (6 mentions) Itaq sybr green master mix, by Bio-Rad (2 mentions) Iscript rt supermix, by Bio-Rad (2 mentions) Cfx connect real time system, by Bio-Rad (2 mentions) Rip lysis buffer, by Solarbio (2 mentions) Genelute total rna purification kit, by Merck Group (2 mentions) Ezh2 antibody, by Abcam (1 mentions) Protease inhibitor, by Thermo Fisher Scientific (1 mentions) Magnetic bead, by Thermo Fisher Scientific (1 mentions) Anti ago2 antibody, by Abcam (1 mentions) Proteinase k, by Absin (1 mentions) Control anti igg antibody, by Abcam (1 mentions) Anti ago2 antibody, by Merck Group (1 mentions) Ab5642, by Abcam (1 mentions) Ab11825, by Abcam (1 mentions) Ab9485, by Abcam (1 mentions) Anti ha magnetic beads, by Thermo Fisher Scientific (1 mentions) Ampliscribe t7 flash biotin rna transcription kit, by Illumina (1 mentions) Ab150077, by Abcam (1 mentions) Magna rip kit, by Merck Group (1 mentions)

Close spelling variants of this product

Imprint® RNA Immunoprecipitation Kit (RIP-12RXN, Imprint RNA Immunoprecipitation (RIP) Kit Imprint® RNA Immunoprecipitation (RIP) Kit (RIP Imprint® RNA RIP kit RIP kit

22 protocols using imprint rip kit

1

Exploring MSTO2P-EZH2 Interaction in HT-29 Cells

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RIP assay was used to explore the interaction between MSTO2P and EZH2 in HT-29 cells using Imprint RIP Kit (Millipore, USA). In brief, cells were lysed in a complete lysis buffer for 30 min at 4 °C. Then, the cell supernatant was incubated with magnetic beads bound with IgG antibody (Abcam, USA) or EZH2 antibody (Abcam, USA) for 4 h at 4 °C. After incubation, the beads were washed and eluted. Then, proteinase K was added to remove protein at 55 °C for 30 min. The input and immunoprecipitated RNAs were isolated to assess MSTO2P expression using RT-qPCR assay.
Guo M, & Zhang X. (2022). LncRNA MSTO2P promotes colorectal cancer progression through epigenetically silencing CDKN1A mediated by EZH2. World Journal of Surgical Oncology, 20, 95.
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2

AGO2-RIP Assay for RNA Detection

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Imprint RIP kit (Millipore, Billerica, MA, USA) was adopted for RIP assay. RIP lysis buffer including protease inhibitor (Thermo Fisher Scientific) was used to lyse MHCC97-H, HCCLM3, and Hep 3B cells. Afterward, magnetic beads (Thermo Fisher Scientific) conjugated with anti-AGO2 antibody (Abcam) or control anti-IgG antibody (Abcam) were added. Proteinase K (Absin, Shanghai, China) was selected to digest the proteins in the immunoprecipitated complex. Finally, quantitative real-time RT-PCR was carried out to detect RNA in the complex. Experiments were conducted three times.
Qin M., Meng Y., Luo C., He S., Qin F., Yin Y., Huang J., Zhao H., Hu J., Deng Z., Qiu Y., Hu G., Pan H., Qin Z., Huang Z, & Yi T. (2021). lncRNA PRR34-AS1 promotes HCC development via modulating Wnt/β-catenin pathway by absorbing miR-296-5p and upregulating E2F2 and SOX12. Molecular Therapy. Nucleic Acids, 25, 37-52.
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3

Circular RNA RIP Assay Protocol

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The RIP assay was conducted using an Imprint® RIP kit (Sigma). The transfected chondrocytes were collected in complete RIP lysis buffer, and cell lysates were interacted with magnetic beads that embraced with anti-Ago2 or anti-IgG at 4°C overnight. Afterward, the samples were incubated with proteinase K to digest protein. Finally, the abundance of circ_0020014 and miR-613 was assessed by RT-qPCR.
Yu Z., Cong F., Zhang W., Song T., Zhang S, & Jiang R. (2022). Circular RNA circ_0020014 contributes to osteoarthritis progression via miR-613/ADAMTS5 axis. Bosnian Journal of Basic Medical Sciences, 22(5), 716-727.
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4

Ago2-RIP Assay for lncRNA

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RIP assay was carried out using Imprint® RIP kit (Sigma, Louis, MO, USA) according to manufacturer’s instruction. For RIP assay, VSMCs were lysed in RIP lysis buffer, followed by incubation with magnetic beads embracing Ago2 (Millipore, Billerica, MA, USA) or IgG (Millipore) antibodies for 24 h at 4°C. The level of RNA enriched by RIP was assessed by RT-qPCR.
Feng L., Liu T., Shi J., Wang Y., Yang Y., Xiao W, & Bai Y. (2023). Circ-UBR4 regulates the proliferation, migration, inflammation, and apoptosis in ox-LDL-induced vascular smooth muscle cells via miR-515-5p/IGF2 axis. Open Medicine, 18(1), 20230751.
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5

RIP Assay for hnRNPK Interacting RNAs

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RIP experiments were carried out with the Imprint RIP Kit (Sigma-Aldrich, St. Louis, USA) using 5 μg of rabbit anti-HnRNPK antibody (ET1610-38, Huabio, Hangzhou, China) or rabbit IgG. The coprecipitated RNAs were detected by qRT‐PCR.
Li T., Wang H., Jiang Y., Chen S., Huang D., Wu Z., Yin X., Zhou C., Li Y, & Zou S. (2023). LITTIP/Lgr6/HnRNPK complex regulates cementogenesis via Wnt signaling. International Journal of Oral Science, 15, 33.
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6

RNA-Immunoprecipitation and RT-qPCR Analysis

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RNA was extracted from RIP reactions and from 0.9% input RNA. Input and RIP RNAs were used for cDNA synthesis with half of the RNA used for mock reverse transcription (no RT enzyme) and the other half used for reverse transcription with the iScript RT Supermix (BioRad). cDNA samples which were then further diluted (1:5 to 1:20). RT-qPCR was performed as described above, using iTaq SYBR Green master mix (BioRad) on the CFX-Connect Real-Time system (BioRad). Relative RIP enrichments were calculated as previously described (Sigma, Imprint® RIP kit): HA RIP Cq values were first compared to IgG control Cq values using the ΔCq method and then normalized across biological replicates by normalizing for input expression levels (1% input, ΔΔCq method). Primer pairs targeting 18s rRNA were used as a non-target control (NT) as well as to assess RIP efficiency by quantification of 18s rRNA de-enrichment between paired input to IP samples. Plots and statistical analyses were generated using the GraphPad Prism (8.2.1) software suite.
Choi S.H., Flamand M.N., Liu B., Zhu H., Hu M., Wang M., Sewell J., Holley C.L., Al-Hashimi H.M, & Meyer K.D. (2022). RBM45 is an m6A-binding protein that affects neuronal differentiation and the splicing of a subset of mRNAs. Cell reports, 40(9), 111293.
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7

Imprint RIP Kit Calculation

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The following calculation was adapted from the Sigma-aldrich Imprint RIP Kit protocol.
Flora P., Wong-Deyrup S.W., Martin E.T., Palumbo R.J., Nasrallah M., Oligney A., Blatt P., Patel D., Fuchs G, & Rangan P. (2018). Sequential Regulation of Maternal mRNAs through a Conserved cis-Acting Element in Their 3′ UTRs. Cell reports, 25(13), 3828-3843.e9.
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8

Investigating miR-203 Binding to HOTAIR lncRNA

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RNA immunoprecipitation (RIP) was performed to investigate the binding of miR-203 to lncRNA HOTAIR. An imprint RIP kit was used as per manufacturer’s instructions (Sigma-Aldrich, St. Louis, MO, USA). Ago2 and IgG (control) antibodies were used for immunoprecipitation. The RIP RNA fraction was reverse transcribed to cDNA using High capacity cDNA reverse transcription kit (Thermo Fisher). Final analysis was performed using RT-qPCR and shown as fold enrichment of HOTAIR to Ago2 with respect to IgG.
Dasgupta P., Kulkarni P., Majid S., Varahram S., Hashimoto Y., Bhat N.S., Shiina M., Deng G., Saini S., Tabatabai Z.L., Yamamura S., Tanaka Y, & Dahiya R. (2018). MicroRNA-203 inhibits long noncoding RNA HOTAIR and regulates tumorigenesis through epithelial-to-mesenchymal transition pathway in renal cell carcinoma. Molecular cancer therapeutics, 17(5), 1061-1069.
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9

miR-107 Binding to HOTAIRM1 Investigated

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RIP assays were performed, using B-CPAP and TPC-1 cell lines, to investigate the binding of miR-107 to HOTAIRM1. An Imprint RIP kit was used according to the manufacturer’s instructions (Sigma-Aldrich), with an anti-Ago2 antibody (Sigma-Aldrich). Total RNA was isolated using a GenElute™ Total RNA Purification Kit (Sigma-Aldrich) and the final analysis was performed using qRT-PCR, as described above.
Li D., Chai L., Yu X., Song Y., Zhu X., Fan S., Jiang W., Qiao T., Tong J., Liu S., Fan L, & Lv Z. (2020). The HOTAIRM1/miR-107/TDG axis regulates papillary thyroid cancer cell proliferation and invasion. Cell Death & Disease, 11(4), 227.
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10

Imprint RIP Kit Calculation

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The following calculation was adapted from the Sigma-aldrich Imprint RIP Kit protocol.
Flora P., Wong-Deyrup S.W., Martin E.T., Palumbo R.J., Nasrallah M., Oligney A., Blatt P., Patel D., Fuchs G, & Rangan P. (2018). Sequential Regulation of Maternal mRNAs through a Conserved cis-Acting Element in Their 3′ UTRs. Cell reports, 25(13), 3828-3843.e9.
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