Anti-phospho-FKHRL1/FOXO3A (Thr32)

Application

Research CategorySignaling

Peptide Inhibition Assay: A 1:500 dilution from a representative lot blocked signal in HEK293 cells treated with IGF.

Immunohistochemistry Analysis: A 1:150 dilution from a representative lot detected phospho-FKHRL1/FOXO3A (Thr32) in human tonsil, human small intestine, and human liver cancer tissues.

Anti-phospho-FKHRL1/FOXO3A (Thr32), Cat. No. ABS554, is a rabbit polyclonal antibody that detects Forkhead box protein O3 and has been tested for use in Immunohistochemistry (Paraffin), Peptide Inhibition Assay, and Western Blotting.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Forkhead box protein O3 (UniProt: O43524; also known as AF6q21 protein, Forkhead in rhabdomyosarcoma-like 1) is encoded by the FOXO3 (also known as FKHRL1, FOXO3A) gene (Gene ID: 2309) in human. Forkhead box protein O3 is a transcriptional activator that riggers apoptosis in the absence of survival factors, including neuronal cell death upon oxidative stress. It recognizes and binds to the DNA sequence 5′-[AG]TAAA[TC]A-3′. Its DNA binding domain located in amino acids 157-251. The nuclear localization signal is located in amino acids 242-259. In the cytoplasm it is present in an inactive state and it translocates to the nucleus upon oxidative stress and in the absence of survival factors. In the presence of survival factors such as IGF-1, it is phosphorylated on Thr32 and Ser253 by AKT1/PKB. The phosphorylated form then interacts with 14-3-3 proteins and is retained in the cytoplasm. Survival factor withdrawal induces dephosphorylation and promotes translocation to the nucleus where the dephosphorylated protein induces transcription of target genes and triggers apoptosis. Forkhead box protein O3 deacetylation by SIRT1 or SIRT2 stimulates its interaction SKP2 and facilitates SCF(SKP2)-mediated FOXO3 ubiquitination and proteasomal degradation. A chromosomal aberration involving FOXO3 is found in secondary acute leukemias.

Immunogen

Epitope: N-terminus

KLH-conjugated linear peptide corresponding to 11 amino acids surrounding phosphorylated Thr32 from the N-terminal region of human Forkhead box protein O3.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Affinity Purified

Quality

Evaluated by Western Blotting in HEK293 treated with IGF stimulation cell lysate.

Western Blotting Analysis: A 1:100 dilution of this antibody detected phospho-FKHRL1/FOXO3A (Thr32) in lysate from HEK293 cells treated with IGF (25 ng/mL for 10 min.).

Specificity

This rabbit polyclonal antibody detects Forkhead box protein O3 in human cells. It targets an epitope within 11 amino acids surrounding phosphorylated Thr32 from the N-terminal region.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Target description

~75 kDa observed; 71.28 kDa calculated. Uncharacterized bands may be observed in some lysate(s).