The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/5000. Detects a band of approximately 90 kDa (predicted molecular weight: 77 kDa).
Use at 2-5 µg/mg of lysate.
Functions as a coactivator for aryl hydrocarbon and nuclear receptors (NR). Recruited to promoters through its contact with the N terminal basic helix-loop-helix-Per-Arnt-Sim (PAS) domain of transcription factors or coactivators, such as NCOA2. During ER-activation acts synergistically in combination with other NCOA2-binding proteins, such as EP300, CREBBP and CARM1. Involved in the transcriptional activation of target genes in the Wnt/CTNNB1 pathway. Functions as a secondary coactivator in LEF1 mediated transcriptional activation via its interaction with CTNNB1. Coactivator function for nuclear receptors and LEF1/CTNNB1 involves differential utilization of two different activation regions.
Cytoplasm. Nucleus. Note: shuttles between nucleus and cytoplasm.
Detection of human CoCoA by Immunoprecipitation in whole cell lysate from MCF7 cells (1 mg, 20% of immunoprecipitate loaded/lane), using ab70564 at 3 µg/mg of lysate (lane 2).
CoCoA was also immunoprecipitated using another CoCoA antibody (ab70563) which recognizes an upstream epitope (lane 1).
Lane 3: Control IgG immunoprecipitate.
Subsequent blotting of immunoprecipitate for the detection of CoCoA was performed using 1µg/ml of ab70564 and 50µg of lysate.
Detection was by chemiluminescence with an exposure time of 30 seconds.