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Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human FOXO3A.
(Peptide available as ab27885.)
Our Abpromise guarantee covers the use of ab31109 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
FOXO3A was immunoprecipitated using 0.5mg PC12 whole cell extract, 5µg of Rabbit polyclonal to FOXO3A and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, PC12 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab31109.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 63kDa; FOXO3A
ab31109 staining FOXO3A in adenomas from ApcMin/+ mice (30 weeks old) by Immunohistochemistry (paraffin embedded sections). Intestines were opened longitudinally, cleaned, Swiss-rolled, fixed with periodate-lysine-paraformaldehyde, and embedded in paraffin.
Phosphorylated FOXO3A was identified utilizing ab31109, followed by HRP-conjugated goat-anti-rabbit IgG. All stains were then developed in 3, 3′-diaminobenzidine and a haematoxylin counterstain was applied.