Cell Signaling Technology
NF-κB p65 (L8F6) Mouse mAb
6956
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10x10^6 cells were resuspended in 500uL of NP40 buffer, incubated on ice for 5 min and spun at 14000g for 10 min. Supernatant was collected and added to 20 ul of protein A/G beads (pre-incubated with 3ug of NF-κB p65 (L8F6) Mouse mAb for 3h at 4'c) and kept at 4'C 0/N. Centriguged and washed the beads for 3 times with lysis buffer. 40uL of 2x laemelli buffer was added and boiled for 10min. Centrifuged and supernatant was loaded on 10% SDS-PAGE
Immunoprecipitation
Glioma stem cells
Used rabbit primary p65 antibody to detect IPed P65
Observed a P65 band at 73 KD which corresponds to P65 protein by western blot, which was not observed in negative controls (antibody with beads, beads with lysate)
N/A
Good antibody for Immunoprecipitation
Good antibody for Immunoprecipitation, Iped P65 runs at a higher molecular weight compared with cell lysate control.