Co-immunoprecipitation
schekman, wusj
Published: 2023-08-26 DOI: 10.17504/protocols.io.14egn32zzl5d/v1
Abstract
This protocol describes a common procedure to perform co-immunoprecipitation
Steps
Co-immunoprecipitation
1.
Media fractions were collected and centrifuged at 1000×g for 0h 10m 0s
2.
The supernatant fractions were collected and concentrated (20-fold) using a 10 kDa Amicon filter.
3.
Concentrated media fractions (1 ml) were incubated with 20 μl of anti-FLAG M2 affinity gel for 1h 0m 0s at 4°C
4.
After washing 5 times with lysis buffer , SDS-PAGE sample loading buffer was added to the beads (twice the beads volume) and samples were processed for SDS-PAGE and immunoblot.
