RIP assay was performed by using the RNA immunoprecipitation kit (GENESEED, China) according to the manufacturer’s instructions. The cell samples were collected and lysed by the lysis buffer containing protease inhibitors and RNase inhibitors. 100 μL supernatant was taken as input control, the negative control was normal rabbit anti-IgG antibody (Cell Signaling Technology, United States). The antibody was linked to the magnetic beads in advance to capture the antigen. Finally the RNA bound to the magnetic beads was eluted and purified for qRT-PCR analysis.
Normal rabbit anti igg antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Normal rabbit anti-IgG antibody is a purified polyclonal antibody raised in rabbits against immunoglobulin G (IgG) from an unspecified source. This antibody can be used as a control or reference antibody in various immunological applications.
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2 protocols using normal rabbit anti igg antibody
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RNA Immunoprecipitation Assay Workflow
2
RNA-binding Protein Immunoprecipitation Assay
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RIP assays were performed using an EZ-Magna RIP™ RNA-binding protein immunoprecipitation kit (Millipore, 17–701) according to the manufacturer’s instructions. Cells at approximately 90% confluence was lysed using complete RIP lysis buffer containing RNase inhibitor and protease inhibitor and then 100 μL of whole cell extract was incubated with RIP buffer containing magnetic beads conjugated to specific antibody (Cell Signaling Technology, 12,582). The nonspecific control was normal rabbit anti-IgG antibody (Cell Signaling Technology, 2729). Purified RNA was subjected to qRT-PCR analysis.
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