To confirm the relationship between circLRIG1, miR-214-3p, and LRIG1 in bladder carcinoma cells, we used the RNA immunoprecipitation assay. Briefly, cells were treated with RIP lysis buffer, and the anti-Ago2 antibody (1:2000; Abcam; USA) or normal IgG antibody (1:2000; Abcam; USA) was conjugated to magnetic beads and incubated with the cell extracts at 4°C for 24 h. The magnetic beads were then harvested and incubated with proteinase K. Finally, we assessed the enrichment of circLRIG1, miR-214-3p, and LRIG1 in the immunoprecipitated RNA by qRT-PCR analysis.
Normal igg antibody
Manufactured by Abcam
Sourced in United States
Normal IgG antibody is a type of immunoglobulin (IgG) that is commonly used as a control in various immunological and biochemical experiments. It serves as a representative of the normal antibody profile found in the body.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using normal igg antibody
1
Investigating circLRIG1-miR-214-3p-LRIG1 Axis
2
RNA Immunoprecipitation Assay for circSCAP
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EZ Magna RIP Kit (Millipore) was used for RNA immunoprecipitation (RIP) following the manufacturer’s instructions. Briefly, cells were lysed in complete RIP lysis buffer. Then the cell lysates were incubated with magnetic beads conjugated with an anti‑Ago2 antibody (1:2000, Abcam, USA) or a normal IgG antibody (1:2000, Abcam, USA) for 24 h at 4 °C. After that, the magnetic beads were washed and treated with proteinase K (50 μg/mL) at 37 °C for 1 h to remove protein. Finally, circSCAP, miR-7, and SMAD2 in immunoprecipitated RNAs were quantified by qRT‐PCR analysis (Qin et al. 2021 (link)).
3
Confirming miR-498 and MDM2 Interaction
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RNA immunoprecipitation assay was utilized to confirm the relationship between miR-498 and MDM2 in BC cells. In short, cells were treated with RIP lysis buffer. The anti‑Ago2 antibody (1: 2000, Abcam, USA) or normal IgG antibody (1:2000, Abcam, USA) was conjugated to magnetic beads and incubated with the cell extract at 4°C for 24 h. Then, the magnetic beads were harvested and incubated with proteinase K. Lastly, the enrichment of miR-498 and MDM2 in immunoprecipitated RNA was assessed with qRT-PCR analysis [40 (link)].
4
Confirming LINC01232-miR-654-3p Interaction in LUSC
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RNA immunoprecipitation assay was used to confirm the relationship between LINC01232 and miR-654-3p in LUSC cells. Briefly, cells were lysed with RIP lysis buffer. Anti-Ago2 antibody (1:2000, Abcam, USA) or normal IgG antibody (1:2000, Abcam, USA) was conjugated to magnetic beads and incubated with the cell lysate for 24 h at 4 ˚C. The magnetic beads were then harvested and treated with proteinase K. Finally, the enrichment of LINC01232 and miR-181a-5p in immunoprecipitated RNAs was assessed by qRT-PCR assay.
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