Synthetic peptide: C-GLNVGNFTGAKQASSQ, with N-terminally added cysteine, conjugated to maleimide activated KLH, corresponding to amino acids 653-668 of Human FOXO3A.
Pig coronary artery endothelial cells were cross-linked for 10 minutes at room temperature by adding 37% formaldehyde to the culture medium. The fixed cells were lysed and the chromatin was isolated. For ChIP, sheared chromatin was incubated with Anti-FOXO3A antibody - ChIP Grade (ab12162). The cells were incubated with wortmannin (30 nM), SB 203580 (10 µM) and SP 600125 (30 µM) for 30 minutes before the addition of CGJ for 30 minutes. n = 4 different experiments. *P<0.05 versus control, #P<0.05 versus CGJ treatment. Endothelial cells were exposed to CGJ for 30 minutes and then chromatin-bound DNA was immunoprecipitated with an antibody against FoxO3a.
ab12162 staining FOXO3A in Rat fibroblast cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.03% Triton X-100 and blocked with 5% serum for 10 minutes at 25°C. Samples were incubated with primary antibody (1/300) for 1 hour at 25°C.
ab12162 staining FOXO3A in human brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with formaldehyde, permeablized with 0.01% Triton-X 100 and blocked with 5% serum for 4 hours at 25°C. The sample was incubated with primary antibody (1/300) at 4°C for 10 hours. An Alexa Fluor® 594-conjugated donkey anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.
ChIP
Related Products
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ChIP Kit (ab500) |
Compatible
Secondaries
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Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) |
Goat Anti-Rabbit IgG H&L (HRP) (ab205718) |
Isotype
control
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Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870) |
Positive
Controls
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Recombinant Human FOXO3A protein (ab114191) |