The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at 2-5 µg/mg of lysate.
1/1000 - 1/5000. Detects a band of approximately 66 kDa (predicted molecular weight: 66 kDa).
1/100 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
NCOA5 is a nuclear receptor coregulator that can have both coactivator and corepressor functions. It interacts with nuclear receptors for steroids (ESR1 and ESR2) independently of the steroid binding domain (AF2) of the ESR receptors, and with the orphan nuclear receptor NR1D2. It Is also involved in the coactivation of nuclear steroid receptors (ER) as well as the corepression of MYC/c myc in response to 17 beta estradiol (E2).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling NCOA5 with ab70831 at 1/200 (1µg/ml). Detection: DAB.
Western blot - NCOA5 antibody (ab70831)
All lanes : Anti-NCOA5 antibody (ab70831) at 0.04 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 3 µg Lane 4 : 293T whole cell lysate at 50 µg
Developed using the ECL technique.
Predicted band size: 66 kDa Observed band size: 66 kDa
Exposure time: 3 minutes
Immunoprecipitation - NCOA5 antibody (ab70831)
Detection of NCOA5 by Western Blot of Immunprecipitate.
ab70831 at 1µg/ml staining NCOA5 in HeLa whole cell lysate immunoprecipitated using ab70831 at 3µg/mg lysate (1 mg/IP; 20% of IP loaded/lane). Detection: Chemiluminescence with exposure time of 30 seconds.