After treatment, cells were harvested and lysed with IP lysis buffer (Meilunbio, China) containing a protease inhibitor cocktail on ice for 30 min, followed by centrifugation for 5 min at 12,000 rpm at 4°C. The antibodies against the target protein were incubated with Protein A/G-Agarose Beads (Invitrogen, USA) for 4 h at 4 °C, mixed with the lysate, and incubated at 4 °C for 16 h. After four washes with PBS, the Co-IP products were collected and analyzed by western blotting.
Ip lysis buffer
Manufactured by Meilun
Sourced in China
IP lysis buffer is a laboratory reagent used for the extraction and purification of proteins from cell samples. It functions to break down cellular membranes and release the contents, including the proteins of interest, into a liquid solution for further analysis or experimentation.
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Lab products found in correlation
2 protocols using ip lysis buffer
1
Protein-Protein Interaction Isolation
2
Immunoprecipitation and Immunoblotting
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Cells were harvested and washed by PBS before lysed with IP lysis buffer (Meilunbio, China) with 1 mM PMSF. Whole cell lysates obtained by centrifugation were incubated with 2 μg of anti-Flag-beads (sigma) at 4 °C. After washing with IP lysis buffer three times, the immunocomplexes were separated by SDS–PAGE. Immunoblotting was performed following standard procedures.
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