Phospho-FOXO3A(Ser253) Antibody (YA6240)
(Synonyms: FOXO3; FKHRL1; FOXO3A; Forkhead box protein O3; AF6q21 protein; Forkhead in rhabdomyosarcoma-like 1)
Based on 1 Customer Validation
Phospho-FOXO3A(Ser253) Antibody (YA6240) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Phospho-FOXO3A(Ser253).
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Host:
Rabbit
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Isotype:
IgG
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Application:
WB, IHC-P, ICC/IF, ELISA
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Reactivity :
Human, Mouse, Rat
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Formulation:
Supplied as a 0.22 μm filtered solution of PBS, 50% glycerol, 0.05% Proclin 300, 0.05% BSA.
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Conjugation:
Non-conjugated
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
Applications
| Application |
IHC-P
IHC-P: Immunohistochemistry-Paraffin
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WB
WB: Western Blot
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ICC/IF
ICC/IF: Immunocytochemistry/
Immunofluorescence |
ELISA
ELISA: Enzyme Linked Immunosorbent Assay
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|---|---|---|---|---|
| Dilution Ratio | 1:200-1:1000 | 1:1000-1:5000 | 1:200-1:1000 | 1:5000-1:20000 |
Product Details
Phospho-FOXO3A(Ser253) Antibody (YA6240) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Phospho-FOXO3A(Ser253).
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Host Rabbit
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Clonality Monoclonal
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Species ReactivityHuman, Mouse, Rat
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Observed Molecular WeightObserved band size: 97 kDa
Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
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Calculated Molecular Weight Predicted band size: 71 kDa
Protein A
Non-conjugated
Phosphorylated
IgG
Product Properties
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Appearance
Solution
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Formulation
Supplied as a 0.22 μm filtered solution of PBS, 50% glycerol, 0.05% Proclin 300, 0.05% BSA.
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Concentration
Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration
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Shipping
Shipping with blue ice.
Verification Images
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Western blot analysis of extracts from Hela (lane 2(20μg), THP-1 (lane 3(20μg), K562 (lane 4(20μg), NIH/3T3 (lane 5(20μg) using Phospho-FOXO3A(Ser253) Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded rat pancreas using Phospho-FOXO3A(Ser253) Antibody (HY-P86548, 1/800). The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using Phospho-FOXO3A(Ser253) Antibody (HY-P86548, 1/500). The section was pretreated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) for 10 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human lymph node tissue using Phospho-FOXO3A(Ser253) Antibody (HY-P86548, 1/500). The section was pretreated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) for 10 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human testis tissue using Phospho-FOXO3A(Ser253) Antibody (HY-P86548, 1/200). The section was pretreated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 15 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human lung tissue using Phospho-FOXO3A(Ser253) Antibody (HY-P86548, 1/200). The section was pretreated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 15 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat spleen tissue using Phospho-FOXO3A(Ser253) Antibody (HY-P86548, 1/200). The section was pretreated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 15 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunocytochemistry analysis of NIH3T3 cells labeling Phospho-FOXO3A (Ser253) Antibody (HY-P86548) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with BSA for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-FOXO3A (Ser253) Antibody (HY-P86548) at 1/200 dilution in BSA for Immunol Staining at 4 ℃ overnight. AF488-conjugated AffiniPure Goat Anti-Rabbit /Mouse IgG H&L（HY-P8002/HY-P8005, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
Background
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Function
FOXO3A transcriptional activator that recognizes and binds to the DNA sequence 5'-[AG]TAAA[TC]A-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[1][2][3][4][5].
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Subcellular Localization
Cytoplasm, cytosol; Nucleus; Mitochondrion matrix; Mitochondrion outer membrane; Peripheral membrane protein; Cytoplasmic side
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Expression
Tissue_specificity:general expression -
Isoforms & Post-Translational Modification
O43524 has 2 isomers: O43524-1: 71277 Da (predicted); O43524-2: 48406 Da (predicted).
In the presence of survival factors such as IGF1, phosphorylated on Thr-32 and Ser-253 by AKT1/PKB (PubMed:10102273). This phosphorylated form then interacts with 14-3-3 proteins and is retained in the cytoplasm (PubMed:10102273). Survival factor withdrawal induces dephosphorylation and promotes translocation to the nucleus where the dephosphorylated protein induces transcription of target genes and triggers apoptosis (PubMed:10102273). Although AKT1/PKB doesn't appear to phosphorylate Ser-315 directly, it may activate other kinases that trigger phosphorylation at this residue (PubMed:10102273, PubMed:11154281). Phosphorylated by STK4/MST1 on Ser-209 upon oxidative stress, which leads to dissociation from YWHAB/14-3-3-beta and nuclear translocation (PubMed:16751106). Phosphorylated by PIM1 (PubMed:18593906). Phosphorylation by AMPK leads to the activation of transcriptional activity without affecting subcellular localization (PubMed:17711846). In response to metabolic stress, phosphorylated by AMPK on Ser-30 which mediates FOXO3 mitochondrial translocation (PubMed:29445193). 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 (PubMed:21329882). Phosphorylated by CHUK/IKKA and IKBKB/IKKB (PubMed:15084260). 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 (PubMed:15084260). May be dephosphorylated by calcineurin A on Ser-299 which abolishes FOXO3 transcriptional activity (By similarity). In cancer cells, ERK mediated-phosphorylation of Ser-12 is required for mitochondrial translocation of FOXO3 in response to metabolic stress or chemotherapeutic agents (PubMed:29445193). Phosphorylation at Ser-253 promotes its degradation by the proteasome (PubMed:30513302). Dephosphorylation at Ser-253 by protein phosphatase 2A (PPP2CA) promotes its stabilization; interaction with PPP2CA is enhanced by AMBRA1 (PubMed:30513302);Deacetylation by SIRT1 or SIRT2 stimulates interaction of FOXO3 with SKP2 and facilitates SCF(SKP2)-mediated FOXO3 ubiquitination and proteasomal degradation (PubMed:21841822). Deacetylation by SIRT2 stimulates FOXO3-mediated transcriptional activity in response to oxidative stress (By similarity). Deacetylated by SIRT3 (PubMed:23283301). Deacetylation by SIRT3 stimulates FOXO3-mediated mtDNA transcriptional activity in response to metabolic stress (PubMed:23283301);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 -
Subunit
Upon metabolic stress, forms a complex composed of FOXO3, SIRT3 and mitochondrial RNA polymerase POLRMT; the complex is recruited to mtDNA in a SIRT3-dependent manner (PubMed:23283301). Also forms a complex composed of FOXO3, SIRT3, TFAM and POLRMT (PubMed:29445193). Interacts with SIRT2; the interaction occurs independently of SIRT2 deacetylase activity (By similarity). Interacts with YWHAB/14-3-3-beta and YWHAZ/14-3-3-zeta, which are required for cytosolic sequestration (PubMed:16751106). Upon oxidative stress, interacts with STK4/MST1, which disrupts interaction with YWHAB/14-3-3-beta and leads to nuclear translocation (PubMed:16751106). Interacts with PIM1 (PubMed:18593906). Interacts with DDIT3/CHOP (PubMed:22761832). Interacts (deacetylated form) with SKP2 (PubMed:21841822). Interacts with CHUK and IKBKB (PubMed:15084260, PubMed:22313691). Interacts with CAMK2A, CAMK2B and calcineurin A (By similarity). Interacts with NUPR1; this interaction represses FOXO3 transactivation (PubMed:20181828)
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SwissProt ID
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Synonyms
FOXO3; FKHRL1; FOXO3A; Forkhead box protein O3; AF6q21 protein; Forkhead in rhabdomyosarcoma-like 1
Documentation
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Data Sheet (235 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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User Guide for Antibodies (1077 KB)
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