Immunoprecipitation kit (ab206996)

Co-immunoprecipitation experiments indicate IKKε does not interact with NIK or IKKε.Immunoprecipitation was performed on MDA MB 468 cells using the Abcam immunoprecipitation kit (ab206996), according to manufacturer's instructions. Briefly, non-denaturing lysis buffer was used to collect 300 µg of cell lysate was incubated overnight with 3 µg/ml of either control rabbit IgG or IKKε rabbit polyclonal antibody (ab7891). Antibody bound proteins were captured using protein A/G sepharose beads, eluted, and analyzed via western blot.

Huh7 cultured with Si-OH, Si-NH2, or PS-NH2 NPs (all 50 µg/ml) bearing BSA or RNase (both 50 µM) as hard protein corona or bare NPs for 4 h. After 4 h post NP treatment, cells were lysed with lysis buffer from immunoprecipitation kit (Abcam). RagC-mTOR complexes were co-immunoprecipitated from the precleared cell lysates with appropriate Ab as described in the manufacturer's instruction. The resulting protein complex was eluted from the beads with Laemmli protein sample buffer for SDS-PAGE and resolved on SDS-PAGE with specific antibody against mTOR

Comparison of immunoprecipitation using Protein A beads and Immunoprecipitation kit ab206996. HDAC2 activity assay of the antigen-antibody complex captured using Protein A beads or ab206996 by following the same protocol demonstrates that ab206996 is more efficient in IP than Protein A beads.