SCC-15 and HSC-3 cells were lysed via RIP lysis buffer (Bio-Rad) at 48 h post transfection. Subsequently, the cell lysate was incubated overnight at 4°C with Protein A/G Magnetic beads (Pierce, Rockford, IL, USA) containing argonaute 2 antibody (anti-Ago2; Abcam, Cambridge, UK) or immunoglobulin G antibody (anti-IgG; Abcam). The RNA enrichments of circGDI2, miR-454-3p and FOXF2 from purified RNA immune-precipitate were tested by qRT-PCR.
Rip lysis buffer
Manufactured by Bio-Rad
Sourced in United States
RIP lysis buffer is a reagent used for the extraction and isolation of ribonucleoprotein (RNP) complexes from cell samples. It is designed to maintain the integrity of RNA-protein interactions during the lysis process.
Automatically generated - may contain errors
3 protocols using rip lysis buffer
1
Profiling circRNAs and miRNAs by RIP-qPCR
2
Argonaute 2-mediated circRNA and miRNA detection
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After transfection for 48 h, HGC27 and AGS cells were lysed using RIP lysis buffer (Bio-Rad, Hercules, CA, USA). Subsequently, cell lysates were incubated with Magnetic beads (Bio-Rad, Hercules, CA, USA) containing Argonaute 2 antibody (Ago2; Bio-Rad, Hercules, CA, USA) or Immunoglobulin G antibody (IgG; Bio-Rad, Hercules, CA, USA) for 3 h at 4℃. The RNA levels of circ_0000620, miR-671-5p and MMP2 were detected by qRT-PCR after RNA extraction using TRIzol reagent (Takara, Dalian, China).
3
Immunoprecipitation of Argonaute 2 in NSCLC
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NSCLC cells were lysed using RIP lysis buffer (Bio-Rad). Protein-A Sepharose beads (Bio-Rad) were precoated with Argonaute 2 antibody (anti-Ago2; Bio-Rad) or immunoglobulin G antibody (anti-IgG; Bio-Rad). The cell lysate was incubated with the precoated Protein-A Sepharose beads for 3 h at 4℃. RNA was extracted using TRIzol reagent (Takara) and detected by RT-qPCR.
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