Co-immunoprecipitation (Co-IP) was carried out using the Pierce Co-IP Kit (Thermo Fisher Scientific, Waltham, USA) based on the manufacturer’s instructions. The total protein in each cell culture was extracted and quantified. A total of 3 mg of protein in 400 μL of supernatant was incubated with 10 μg of anti-NAP1L1, anti-HDGF (Proteintech), anti-DDX5 (Abcam), or anti-IgG antibodies (Proteintech) for 12 h at 4°C. The beads were then washed, eluted in a sample buffer, and boiled for 10 min at 100°C. The immune complexes were then subjected to western blot analysis, with anti-IgG used as a negative control.
Anti igg antibodies
Manufactured by Proteintech
Anti-IgG antibodies are laboratory reagents used to detect and analyze immunoglobulin G (IgG) proteins. These antibodies specifically bind to IgG molecules, allowing for their identification and quantification in various experimental and diagnostic applications.
Automatically generated - may contain errors
2 protocols using anti igg antibodies
1
Co-immunoprecipitation Analysis of Protein Complexes
2
Immunoprecipitation and Western Blotting
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The cells were collected and cleaved in low-salt lysis buffer for 30 min at 4°C. The samples were incubated with immunoglobulin microspheres at 4°C for 2 hours and then centrifuged at 5000 rpm for 10 minutes. The supernatant (20 μl) was saved as “Input”. Anti-IgG antibodies (Proteintech, Wuhan, China) or anti-Flag antibodies (Proteintech, Wuhan, China) were added to the remaining cell lysate and prewashed with TBST buffer 3 times. The cell lysate with antibody was rotated gently overnight at 4°C. The protein A/G microspheres were added to the cell lysate and then rotated at 4°C for 4 hours to harvest microbeads. The beads were washed in TBST 3 times and then boiled for 10 min in 2X sodium dodecyl sulfate loading buffer. The follow-up analysis was performed by Western blotting.
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