• Immunohistochemistry analysis of paraffin-embedded human breast carcinoma, using FKHRL1 (Phospho-Ser253) Antibody. The picture on the right is blocked with the phospho peptide.
  • Western blot analysis of lysates from LOVO cells treated with serum 20% 30', using FKHRL1 (Phospho-Ser253) Antibody. The lane on the right is blocked with the phospho peptide.
  • Western blot analysis of HELA cells using Phospho-FoxO3A (S253) Polyclonal Antibody diluted at 1:2000
  • Immunohistochemical analysis of paraffin-embedded Human brain. Antibody was diluted at 1:100 (4°C overnight). High-pressure and temperature Tris-EDTA, pH8.0 was used for antigen retrieval. Negetive contrl (right) obtaned from antibody was pre-absorbed by immunogen peptide.
  • Immunohistochemical analysis of paraffin-embedded Mouse-liver tissue. 1, FoxO3A (phospho Ser253) Polyclonal Antibody was diluted at 1:200 (4°C, overnight). 2, Sodium citrate pH 6.0 was used for antibody retrieval (>98°C, 20min). 3, Secondary antibody was diluted at 1:200 (room tempeRature, 30min). Negative control was used by secondary antibody only.
  • Immunohistochemical analysis of paraffin-embedded Mouse-colon tissue. 1, FoxO3A (phospho Ser253) Polyclonal Antibody was diluted at 1:200 (4°C, overnight). 2, Sodium citrate pH 6.0 was used for antibody retrieval (>98°C, 20min). 3, Secondary antibody was diluted at 1:200 (room tempeRature, 30min). Negative control was used by secondary antibody only.
  • Immunohistochemical analysis of paraffin-embedded Mouse-testis tissue. 1, FoxO3A (phospho Ser253) Polyclonal Antibody was diluted at 1:200 (4°C, overnight). 2, Sodium citrate pH 6.0 was used for antibody retrieval (>98°C, 20min). 3, Secondary antibody was diluted at 1:200 (room tempeRature, 30min). Negative control was used by secondary antibody only.
  • Immunohistochemical analysis of paraffin-embedded Rat-spleen tissue. 1, FoxO3A (phospho Ser253) Polyclonal Antibody was diluted at 1:200 (4°C, overnight). 2, Sodium citrate pH 6.0 was used for antibody retrieval (>98°C, 20min). 3, Secondary antibody was diluted at 1:200 (room tempeRature, 30min). Negative control was used by secondary antibody only.
  • Immunohistochemical analysis of paraffin-embedded Rat-lung tissue. 1, FoxO3A (phospho Ser253) Polyclonal Antibody was diluted at 1:200 (4°C, overnight). 2, Sodium citrate pH 6.0 was used for antibody retrieval (>98°C, 20min). 3, Secondary antibody was diluted at 1:200 (room tempeRature, 30min). Negative control was used by secondary antibody only.
  • Immunohistochemical analysis of paraffin-embedded Rat-testis tissue. 1, FoxO3A (phospho Ser253) Polyclonal Antibody was diluted at 1:200 (4°C, overnight). 2, Sodium citrate pH 6.0 was used for antibody retrieval (>98°C, 20min). 3, Secondary antibody was diluted at 1:200 (room tempeRature, 30min). Negative control was used by secondary antibody only.
  • Immunohistochemical analysis of paraffin-embedded Human-colon tissue. 1, FoxO3A (phospho Ser253) Polyclonal Antibody was diluted at 1:200 (4°C, overnight). 2, Sodium citrate pH 6.0 was used for antibody retrieval (>98°C, 20min). 3, Secondary antibody was diluted at 1:200 (room tempeRature, 30min). Negative control was used by secondary antibody only.

Anti-Phospho-FOXO3-Ser253 antibody (220-269 aa) (STJ90275)

SKU:
STJ90275

Current Stock:
Host: Rabbit
Applications: WB/IHC/IF/ELISA
Reactivity: Human/Mouse/Rat/Zebrafish
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-Forkhead box protein O3-Ser253 (220-269 aa) is suitable for use in Western Blot, Immunohistochemistry, Immunofluorescence and ELISA research applications.
Clonality: Polyclonal
Conjugation: Unconjugated
Isotype: IgG
Formulation: Liquid in PBS containing 50% Glycerol, 0.5% BSA and 0.02% Sodium Azide.
Purification: The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
Concentration: 1 mg/mL
Dilution Range: WB 1:500-1:2000
IHC 1:100-1:300
IF 1:200-1:1000
ELISA 1:20000
Storage Instruction: Store at-20°C for up to 1 year from the date of receipt, and avoid repeat freeze-thaw cycles.
Gene Symbol: FOXO3
Gene ID: 2309
Uniprot ID: FOXO3_HUMAN
Immunogen Region: 220-269 aa
Specificity: Phospho-FoxO3A (S253) Polyclonal Antibody detects endogenous levels of FoxO3A protein only when phosphorylated at S253.
Immunogen: The antiserum was produced against synthesized peptide derived from the human FKHRL1 around the phosphorylation site of Ser253 at the amino acid range 220-269
Post Translational Modifications In the presence of survival factors such as IGF-1, phosphorylated on Thr-32 and Ser-253 by AKT1/PKB. This 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. Although AKT1/PKB doesn't appear to phosphorylate Ser-315 directly, it may activate other kinases that trigger phosphorylation at this residue. Phosphorylated by STK4/MST1 on Ser-209 upon oxidative stress, which leads to dissociation from YWHAB/14-3-3-beta and nuclear translocation. Phosphorylated by PIM1. Phosphorylation by AMPK leads to the activation of transcriptional activity without affecting subcellular localization. In response to metabolic stress, phosphorylated by AMPK on Ser-30 which mediates FOXO3 mitochondrial translocation. Phosphorylation by MAPKAPK5 promotes nuclear localization and DNA-binding, leading to induction of miR-34b and miR-34c expression, 2 post-transcriptional regulators of MYC that bind to the 3'UTR of MYC transcript and prevent its translation. Phosphorylated by CHUK/IKKA and IKBKB/IKKB. TNF-induced inactivation of FOXO3 requires its phosphorylation at Ser-644 by IKBKB/IKKB which promotes FOXO3 retention in the cytoplasm, polyubiquitination and ubiquitin-mediated proteasomal degradation. May be dephosphorylated by calcineurin A on Ser-299 which abolishes FOXO3 transcriptional activity. In cancer cells, ERK mediated-phosphorylation of Ser-12 is required for mitochondrial translocation of FOXO3 in response to metabolic stress or chemotherapeutic agents. Phosphorylation at Ser-253 promotes its degradation by the proteasome. Dephosphorylation at Ser-253 by protein phosphatase 2A (PPP2CA) promotes its stabilization.interaction with PPP2CA is enhanced by AMBRA1. Deacetylation by SIRT1 or SIRT2 stimulates interaction of FOXO3 with SKP2 and facilitates SCF(SKP2)-mediated FOXO3 ubiquitination and proteasomal degradation. Deacetylation by SIRT2 stimulates FOXO3-mediated transcriptional activity in response to oxidative stress. Deacetylated by SIRT3. Deacetylation by SIRT3 stimulates FOXO3-mediated mtDNA transcriptional activity in response to metabolic stress. Heavily methylated by SET9 which decreases stability, while moderately increasing transcriptional activity. The main methylation site is Lys-271. Methylation doesn't affect subcellular location. Polyubiquitinated. Ubiquitinated by a SCF complex containing SKP2, leading to proteasomal degradation. The N-terminus is cleaved following import into the mitochondrion.
Function Transcriptional activator that recognizes and binds to the DNA sequence 5'-AGTAAATCA-3' and regulates different processes, such as apoptosis and autophagy. Acts as a positive regulator of autophagy in skeletal muscle: in starved cells, 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. Triggers apoptosis in the absence of survival factors, including neuronal cell death upon oxidative stress. Participates in post-transcriptional regulation of MYC: following phosphorylation by MAPKAPK5, promotes induction of miR-34b and miR-34c expression, 2 post-transcriptional regulators of MYC that bind to the 3'UTR of MYC transcript and prevent its translation. In response to metabolic stress, translocates into the mitochondria where it promotes mtDNA transcription. In response to metabolic stress, translocates into the mitochondria where it promotes mtDNA transcription. 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. Also acts as a key regulator of regulatory T-cells (Treg) differentiation by activating expression of FOXP3.
Protein Name Forkhead Box Protein O3
Af6q21 Protein
Forkhead In Rhabdomyosarcoma-Like 1
Database Links Reactome: R-HSA-1181150
Reactome: R-HSA-198693
Reactome: R-HSA-5674400
Reactome: R-HSA-5687128
Reactome: R-HSA-6785807
Reactome: R-HSA-8862803
Reactome: R-HSA-8952158
Reactome: R-HSA-9607240
Reactome: R-HSA-9614399
Reactome: R-HSA-9614657
Reactome: R-HSA-9615017
Reactome: R-HSA-9617629
Reactome: R-HSA-9617828
Reactome: R-HSA-9634638
Cellular Localisation Cytoplasm
Cytosol
Nucleus
Mitochondrion Matrix
Mitochondrion Outer Membrane
Peripheral Membrane Protein
Cytoplasmic Side
Retention In The Cytoplasm Contributes To Its Inactivation
Translocates To The Nucleus Upon Oxidative Stress And In The Absence Of Survival Factors
Translocates From The Cytosol To The Nucleus Following Dephosphorylation In Response To Autophagy-Inducing Stimuli
Translocates In A Ampk-Dependent Manner Into The Mitochondrion In Response To Metabolic Stress
Serum Deprivation Increases Localization To The Nucleus
Leading To Activate Expression Of Sox9 And Subsequent Chondrogenesis
Alternative Antibody Names Anti-Forkhead Box Protein O3 antibody
Anti-Af6q21 Protein antibody
Anti-Forkhead In Rhabdomyosarcoma-Like 1 antibody
Anti-FOXO3 antibody
Anti-FKHRL1 antibody
Anti-FOXO3A antibody

Information sourced from Uniprot.org

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