Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Osteosarcoma)
Total protein in input
500 µg
Treatment
Exogenous expression of GFP-FOXM1 fusion protein. Protein crosslinking with 25 µM DSP for 45 minutes.
Immuno-precipitation step
Protein A
Specification
Osteosarcoma
Other product details
Incubation time
4 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: Pierce IP Lysis Buffer (Thermo 87788)
Concentration
6 µg/mg lysate
Additional data
Additional Notes
FOXM1 (ab207298, EPR17379) antibody was pre-bound to Protein A agarose beads and used for FOXM1 immunoprecipitation. Whole cell lysates were obtained from U2OS cells overexpressing GFP-FOXM1 fusion protein in native conditions or after protein crosslinking with 25 µM DSP. Control rabbit IgG were used for negative control IP samples. EPR17379 antibody recognizes both endogenous FOXM1 protein and GFP-FOXM1 fusion protein. Protein crosslinking reduces overall sensitivity, but the antibody still recognizes and binds FOXM1, including cross-linked FOXM1 dimers. Some FOXM1 partner proteins were co-immunoprecipitated using this antibody, however, I can't show the pictures before they get published. Western Blot samples were prepaired by boiling beads with immunoprecipitated proteins in Laemmli SDS sample buffer with 2-mercaptoethanol for 10 minutes at 100 degrees.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
DR. Mikhail Chesnokov
Verified customer
提交于 Mar 31 2020