Product Name:
FOXO3-pS253
Product Number:
ab-pn821
Target Full Name: Forkhead box protein O3
Target Alias: AF6q21; AF6q21 protein; Fkhr2; Fkhrl1; Forkhead box O3; Forkhead box O3A; Forkhead in rhabdomyosarcoma-like 1; FOXO2; Foxo3; Foxo3a
Product Type Specific: FOXO3 phosphosite-specific antibody
Antibody Code: PN821
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S253
Protein UniProt: O43524
Protein SigNET: FOXO3
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human FOXO3
Antibody Immunogen Sequence: RAV(pS)MDNSC
Antibody Immunogen Description: Corresponds to amino acid residues R250 to S256. S253 phosphorylation inhibits activity, although there is one report of activation and induced interaction with 14-3-3-beta, 14-3-3-epsilon and 14-3-3-zeta. This is one of the major phosphosites with ≥93 mass spectrometry reports recorded in PhosphoSitePlus. FOXO3 is known to be phosphorylated at this site in vitro by Akt1 (PKBa).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 0.9 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Rhesus Macaque | Dog | Rat | Mouse | Frog.
Related Product 1: FOXK1 S441+S445 phosphorylation antibody (Cat. No.: AB-PN590)
Related Product 2: FOXK2 S424+S428 phosphorylation antibody (Cat. No.: AB-PN591)
Related Product 3: FOXM1 T611 phosphorylation antibody (Cat. No.: AB-PN592)
Related Product 4: FOXO1 S256 phosphorylation antibody (Cat. No.: AB-PN593)
Related Product 5: FOXO1 S319 phosphorylation antibody (Cat. No.: AB-PN594)
Related Product 6: FOXO1 S329 phosphorylation antibody (Cat. No.: AB-PN595)
Related Product 7: FOXO3 T32 phosphorylation antibody (Cat. No.: AB-PN596)
Related Product 8: FOXO3 S294 phosphorylation antibody (Cat. No.: AB-PN597)
Scientific Background: FOXO3A (Forkhead box protein O3) is a ubiquitous transcription factor and member of the FOXO family that acts as a master regulator of cellular longevity, stress resistance, and homeostasis. It recognizes and binds to the DNA sequence 5'-[AG]TAAA[TC]A-3' and regulates different processes, such as apoptosis and autophagy. It acts as a positive regulator of autophagy in skeletal muscle. In starved cells, it enters the nucleus following dephosphorylation and binds the promoters of autophagy genes, such as GABARAP1L, MAP1LC3B and ATG12, thereby activating their expression, resulting in proteolysis of skeletal muscle proteins. It triggers apoptosis in the absence of survival factors, including neuronal cell death upon oxidative stress. It participates in post-transcriptional regulation of MYC: following phosphorylation by MAPKAPK5, promotes induction of miR-34b and miR-34c expression, which are two post-transcriptional regulators of MYC that bind to the 3'UTR of MYC transcript and prevent its translation. In response to metabolic stress, FOXO3 translocates into the mitochondria where it promotes mtDNA transcription. FOXO3 activity is controlled by post-translational modifications. Under nutrient-rich conditions, AKT and ERK kinases phosphorylate FOXO3, forcing it out of the nucleus and inhibiting its action. In contrast, SIRT1, AMPK, and conditions like caloric restriction or exercise activate FOXO3. This description may include information annotated by UniProt and/or Google AI.