• Western blot analysis of extracts of 293T cells, using Phospho-FoxO1-S256 antibody (STJ22144). 293T were treated by CIP (20ul CIP for each 400ul cell lysate) at 37 °C for 1 hour. Secondary antibody: HRP Goat Anti-rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% BSA. Detection: ECL Basic Kit. Exposure time: 90s.
  • Western blot analysis of extracts of HeLa cells, using Phospho-FoxO1-S256 polyclonal antibody (STJ22144) at 1:1000 dilution or FOXO1 antibody (STJ115801). Hela cell lysate were treated by CIP (20ul CIP for each 400ul cell lysate) at 37 °C for 1 hour or treated by 10% FBS at 37 °C for 30 minutes after serum-starvation overnight.. Secondary antibody: HRP Goat Anti-rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% BSA. Detection: ECL Basic Kit. Exposure time: 2s.
  • Immunofluorescence analysis of NIH/3T3 using Phospho-FoxO1-S256 antibody (STJ22144) at dilution of 1:50. Blue: DAPI for nuclear staining.
  • Immunofluorescence analysis of PC-12 using Phospho-FoxO1-S256 antibody (STJ22144) at dilution of 1:50. Blue: DAPI for nuclear staining.

Anti-Phospho-FOXO1-S256 antibody (STJ22144)

SKU:
STJ22144

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Host: Rabbit
Applications: WB/IF
Reactivity: Human/Mouse/Rat
Note: STRICTLY FOR FURTHER SCIENTIFIC RESEARCH USE ONLY (RUO). MUST NOT TO BE USED IN DIAGNOSTIC OR THERAPEUTIC APPLICATIONS.
Short Description: Rabbit polyclonal antibody anti-Phospho-FoxO1-S256 is suitable for use in Western Blot and Immunofluorescence research applications.
Clonality: Polyclonal
Conjugation: Unconjugated
Isotype: IgG
Formulation: PBS with 0.01% Thimerosal, 50% Glycerol, pH7.3.
Purification: Affinity purification
Dilution Range: WB 1:500-1:1000
IF/ICC 1:50-1:200
Storage Instruction: Store at-20°C for up to 1 year from the date of receipt, and avoid repeat freeze-thaw cycles.
Gene Symbol: FOXO1
Gene ID: 2308
Uniprot ID: FOXO1_HUMAN
Immunogen: A synthetic phosphorylated peptide around S256 of human FoxO1 (NP_002006.2).
Immunogen Sequence: AASMD
Tissue Specificity Ubiquitous.
Post Translational Modifications Phosphorylation by NLK promotes nuclear export and inhibits the transcriptional activity. In response to growth factors, phosphorylation on Thr-24, Ser-256 and Ser-322 by PKB/AKT1 promotes nuclear export and inactivation of transactivational activity. Phosphorylation on Thr-24 is required for binding 14-3-3 proteins. Phosphorylation of Ser-256 decreases DNA-binding activity and promotes the phosphorylation of Thr-24 and Ser-319, permitting phosphorylation of Ser-322 and Ser-325, probably by CDK1, leading to nuclear exclusion and loss of function. Stress signals, such as response to oxygen or nitric oxide, attenuate the PKB/AKT1-mediated phosphorylation leading to nuclear retention. Phosphorylation of Ser-329 is independent of IGF1 and leads to reduced function. Dephosphorylated on Thr-24 and Ser-256 by PP2A in beta-cells under oxidative stress leading to nuclear retention. Phosphorylation of Ser-249 by CDK1 disrupts binding of 14-3-3 proteins leading to nuclear accumulation and has no effect on DNA-binding nor transcriptional activity. Phosphorylation by STK4/MST1 on Ser-212, upon oxidative stress, inhibits binding to 14-3-3 proteins and nuclear export. PPIA/CYPA promotes its dephosphorylation on Ser-256. Ubiquitinated by SKP2. Ubiquitination leads to proteasomal degradation. Methylation inhibits AKT1-mediated phosphorylation at Ser-256 and is increased by oxidative stress. Acetylated. Acetylation at Lys-262, Lys-265 and Lys-274 are necessary for autophagic cell death induction. Deacetylated by SIRT2 in response to oxidative stress or serum deprivation, thereby negatively regulating FOXO1-mediated autophagic cell death. Once in the nucleus, acetylated by CREBBP/EP300. Acetylation diminishes the interaction with target DNA and attenuates the transcriptional activity. It increases the phosphorylation at Ser-256. Deacetylation by SIRT1 results in reactivation of the transcriptional activity. Oxidative stress by hydrogen peroxide treatment appears to promote deacetylation and uncoupling of insulin-induced phosphorylation. By contrast, resveratrol acts independently of acetylation. Acetylated at Lys-423, promoting its localization to the nucleus and transcription factor activity. Deacetylation at Lys-423 by SIRT6, promotes its translocation into the cytoplasm, preventing its transcription factor activity. Deacetylation and subsequent inhibition by SIRT6 has different effects depending on cell types: it inhibits gluconeogenesis in hepatocytes, promotes glucose sensing in pancreatic beta-cells and regulates lipid catabolism in brown adipocytes.
Function Transcription factor that is the main target of insulin signaling and regulates metabolic homeostasis in response to oxidative stress. Binds to the insulin response element (IRE) with consensus sequence 5'-TTG/ATTTTG-3' and the related Daf-16 family binding element (DBE) with consensus sequence 5'-TTG/ATTTAC-3'. Activity suppressed by insulin. Main regulator of redox balance and osteoblast numbers and controls bone mass. Orchestrates the endocrine function of the skeleton in regulating glucose metabolism. Also acts as a key regulator of chondrogenic commitment of skeletal progenitor cells in response to lipid availability: when lipids levels are low, translocates to the nucleus and promotes expression of SOX9, which induces chondrogenic commitment and suppresses fatty acid oxidation. Acts synergistically with ATF4 to suppress osteocalcin/BGLAP activity, increasing glucose levels and triggering glucose intolerance and insulin insensitivity. Also suppresses the transcriptional activity of RUNX2, an upstream activator of osteocalcin/BGLAP. Acts as an inhibitor of glucose sensing in pancreatic beta cells by acting as a transcription repressor and suppressing expression of PDX1. In hepatocytes, promotes gluconeogenesis by acting together with PPARGC1A and CEBPA to activate the expression of genes such as IGFBP1, G6PC1 and PCK1. Also promotes gluconeogenesis by directly promoting expression of PPARGC1A and G6PC1. Important regulator of cell death acting downstream of CDK1, PKB/AKT1 and STK4/MST1. Promotes neural cell death. Mediates insulin action on adipose tissue. Regulates the expression of adipogenic genes such as PPARG during preadipocyte differentiation and, adipocyte size and adipose tissue-specific gene expression in response to excessive calorie intake. Regulates the transcriptional activity of GADD45A and repair of nitric oxide-damaged DNA in beta-cells. Required for the autophagic cell death induction in response to starvation or oxidative stress in a transcription-independent manner. Mediates the function of MLIP in cardiomyocytes hypertrophy and cardiac remodeling. Regulates endothelial cell (EC) viability and apoptosis in a PPIA/CYPA-dependent manner via transcription of CCL2 and BCL2L11 which are involved in EC chemotaxis and apoptosis.
Protein Name Forkhead Box Protein O1
Forkhead Box Protein O1a
Forkhead In Rhabdomyosarcoma
Database Links Reactome: R-HSA-198693
Reactome: R-HSA-210745
Reactome: R-HSA-211163
Reactome: R-HSA-5674400
Reactome: R-HSA-5687128
Reactome: R-HSA-6785807
Reactome: R-HSA-9614399
Reactome: R-HSA-9614657
Reactome: R-HSA-9615017
Reactome: R-HSA-9617629
Reactome: R-HSA-9617828
Cellular Localisation Cytoplasm
Nucleus
Shuttles Between The Cytoplasm And Nucleus
Largely Nuclear In Unstimulated Cells
In Osteoblasts
Colocalizes With Atf4 And Runx2 In The Nucleus
Serum Deprivation Increases Localization To The Nucleus
Leading To Activate Expression Of Sox9 And Subsequent Chondrogenesis
Insulin-Induced Phosphorylation At Ser-256 By Pkb/Akt1 Leads
Via Stimulation Of Thr-24 Phosphorylation
To Binding Of 14-3-3 Proteins And Nuclear Export To The Cytoplasm Where It Is Degraded By The Ubiquitin-Proteasomal Pathway
Phosphorylation At Ser-249 By Cdk1 Disrupts Binding Of 14-3-3 Proteins And Promotes Nuclear Accumulation
Phosphorylation By Nlk Results In Nuclear Export
Translocates To The Nucleus Upon Oxidative Stress-Induced Phosphorylation At Ser-212 By Stk4/Mst1
Sgk1-Mediated Phosphorylation Also Results In Nuclear Translocation
Retained In The Nucleus Under Stress Stimuli Including Oxidative Stress
Nutrient Deprivation Or Nitric Oxide
Retained In The Nucleus On Methylation
Ppia/Cypa Stimulates Its Nuclear Accumulation
Deacetylation By Sirt6
Promotes Its Translocation Into The Cytoplasm
Alternative Antibody Names Anti-Forkhead Box Protein O1 antibody
Anti-Forkhead Box Protein O1a antibody
Anti-Forkhead In Rhabdomyosarcoma antibody
Anti-FOXO1 antibody
Anti-FKHR antibody
Anti-FOXO1A antibody

Information sourced from Uniprot.org

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