RIP experiment was performed to verify the relationship between LEGLTBC and miR-34a using the EZ-Magna RIP Kit (Millipore, USA). Briefly, the INS-1 cells were lysed in complete RIP lysis buffer containing a protease inhibitor and an RNase inhibitor. Then, 100 μL extract was incubated with argonaute 2 (AGO2) antibody or control immunoglobulin G (IgG; Millipore, USA) for 8 h at 4°C, followed by adding protein A/G beads. Next, the beads were incubated with proteinase K to digest the proteins. Finally, purified RNA was subjected to qRT-PCR analysis using specific primers for LEGLTBC.
Control immunoglobulin g igg
Manufactured by Merck Group
Sourced in United States
Control immunoglobulin G (IgG) is a laboratory reagent used to validate and calibrate immunoassay tests. It serves as a reference material to ensure the accuracy and precision of immunoglobulin G measurements in clinical and research samples.
Automatically generated - may contain errors
Lab products found in correlation
Ez magna rip kit, by Merck Group (1 mentions)
Rip lysis buffer, by Beyotime (1 mentions)
Magna rna rip kit, by Merck Group (1 mentions)
Argonaute2, by Merck Group (1 mentions)
Proteinase k, by Merck Group (1 mentions)
Anti ezh2, by Merck Group (1 mentions)
Magna rip rna binding protein immunoprecipitation kit, by Merck Group (1 mentions)
Anti suz12, by Merck Group (1 mentions)
3 protocols using control immunoglobulin g igg
1
Investigating LEGLTBC and miR-34a Interaction
2
Argonaute-2 Immunoprecipitation for RNA Binding
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RNA binding protein immunoprecipitation (RIP) assay was executed by using Magna RNA RIP Kit (Merck KGaA, Darmstadt, Germany) by following the manufacturer’s protocol. INS-1E cells were lysed by using RIP lysis buffer (Beyotime) and incubated with magnetic beads conjugated with Argonaute-2 (Ago2, Millipore, MA, USA) or control immunoglobulin G (IgG, Millipore) at 4ʹC overnight. Detachment of the complex was with proteinase K (Sigma-Aldrich), and the bound RNA was identified by RT-qPCR [14 (link)].
3
EZH2 and SUZ12 RIP Profiling
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RIP experiments were performed using a Magna RIP RNA-binding protein immunoprecipitation kit (Millipore, Billerica, MA, USA), following the manufacturer's instructions. In brief, PC-9/GR cells were scraped off the culture plate and then lysed in RIP lysis buffer. Cell extract was incubated with RIP buffer containing magnetic beads conjugated with anti-EZH2, anti-SUZ12, or control immunoglobulin G (IgG) (Millipore). Finally, immunoprecipitated RNA was isolated and analyzed by quantitative real-time PCR.
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