Product: Phospho-FOXO1A (Ser256) Antibody
Catalog: AF3417
Description: Rabbit polyclonal antibody to Phospho-FOXO1A (Ser256)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Dog, Chicken, Xenopus
Mol.Wt.: 69kDa; 70kD(Calculated).
Uniprot: Q12778
RRID: AB_2834859

View similar products>>

   Size Price Inventory
 100ul $280 In stock
 200ul $350 In stock

Lead Time: Same day delivery

For pricing and ordering contact:
Local distributors

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Dog(100%), Chicken(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
Phospho-FOXO1A (Ser256) Antibody detects endogenous levels of FOXO1A only when phosphorylated at Serine 256.
RRID:
AB_2834859
Cite Format: Affinity Biosciences Cat# AF3417, RRID:AB_2834859.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

FKH 1; FKH1; FKHR; Forkhead (Drosophila) homolog 1 (rhabdomyosarcoma); Forkhead box O1; Forkhead box protein O1; Forkhead box protein O1A; Forkhead in rhabdomyosarcoma; Forkhead, Drosophila, homolog of, in rhabdomyosarcoma; FoxO transcription factor; foxo1; FOXO1_HUMAN; FOXO1A; OTTHUMP00000018301;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
Q12778 FOXO1_HUMAN:

Ubiquitous.

Description:
This gene belongs to the forkhead family of transcription factors which are characterized by a distinct forkhead domain. The specific function of this gene has not yet been determined; however, it may play a role in myogenic growth and differentiation.
Sequence:
MAEAPQVVEIDPDFEPLPRPRSCTWPLPRPEFSQSNSATSSPAPSGSAAANPDAAAGLPSASAAAVSADFMSNLSLLEESEDFPQAPGSVAAAVAAAAAAAATGGLCGDFQGPEAGCLHPAPPQPPPPGPLSQHPPVPPAAAGPLAGQPRKSSSSRRNAWGNLSYADLITKAIESSAEKRLTLSQIYEWMVKSVPYFKDKGDSNSSAGWKNSIRHNLSLHSKFIRVQNEGTGKSSWWMLNPEGGKSGKSPRRRAASMDNNSKFAKSRSRAAKKKASLQSGQEGAGDSPGSQFSKWPASPGSHSNDDFDNWSTFRPRTSSNASTISGRLSPIMTEQDDLGEGDVHSMVYPPSAAKMASTLPSLSEISNPENMENLLDNLNLLSSPTSLTVSTQSSPGTMMQQTPCYSFAPPNTSLNSPSPNYQKYTYGQSSMSPLPQMPIQTLQDNKSSYGGMSQYNCAPGLLKELLTSDSPPHNDIMTPVDPGVAQPNSRVLGQNVMMGPNSVMSTYGSQASHNKMMNPSSHTHPGHAQQTSAVNGRPLPHTVSTMPHTSGMNRLTQVKTPVQVPLPHPMQMSALGGYSSVSSCNGYGRMGLLHQEKLPSDLDGMFIERLDCDMESIIRNDLMDGDTLDFNFDNVLPNQSFPHSVKTTTHSWVSG

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Bovine
100
Dog
100
Xenopus
100
Chicken
100
Horse
0
Sheep
0
Zebrafish
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q12778 As Substrate

Site PTM Type Enzyme
T24 Phosphorylation P31751 (AKT2) , P31749 (AKT1) , P11309-2 (PIM1) , PR:P31749 (hAKT1)
S152 Phosphorylation
S153 Phosphorylation
S164 Phosphorylation
T182 Phosphorylation
S184 Phosphorylation
S205 Phosphorylation
K210 Ubiquitination
S212 Phosphorylation Q13043 (STK4)
S215 Phosphorylation
S218 Phosphorylation
K222 Acetylation
S234 Phosphorylation
S235 Phosphorylation
K245 Acetylation
S246 Phosphorylation P28482 (MAPK1)
K248 Acetylation
S249 Phosphorylation P06493 (CDK1) , Q00535 (CDK5) , P24941 (CDK2) , P11802 (CDK4)
R251 Methylation
R253 Methylation
S256 Phosphorylation PR:P31749 (hAKT1) , Q13153 (PAK1) , Q16512 (PKN1) , P31749 (AKT1) , P31751 (AKT2) , P11309-2 (PIM1)
K262 Acetylation
K265 Acetylation
K274 Acetylation
S276 Phosphorylation
S287 Phosphorylation
S293 Phosphorylation
K294 Acetylation
S298 Phosphorylation
S301 Phosphorylation
S303 Phosphorylation
T317 O-Glycosylation
S318 O-Glycosylation
S319 Phosphorylation PR:P31749 (hAKT1) , Q13237 (PRKG2) , P31749 (AKT1) , P11309-2 (PIM1)
S322 Phosphorylation P48729 (CSNK1A1) , Q9HCP0 (CSNK1G1)
T323 Phosphorylation
S325 Phosphorylation P48729 (CSNK1A1) , P49841 (GSK3B)
S329 Phosphorylation Q9UBE8 (NLK) , Q13627 (DYRK1A)
T333 Phosphorylation
S383 Phosphorylation
S394 Phosphorylation
T402 Phosphorylation
S413 Phosphorylation P28482 (MAPK1)
S416 Phosphorylation Q16539 (MAPK14)
S418 Phosphorylation P28482 (MAPK1)
S429 Phosphorylation P28482 (MAPK1)
S430 Phosphorylation
S432 Phosphorylation Q16539 (MAPK14)
T467 Phosphorylation
S470 Phosphorylation P28482 (MAPK1) , Q16539 (MAPK14)
T478 Phosphorylation Q16539 (MAPK14) , P28482 (MAPK1)
S505 Phosphorylation
S509 Phosphorylation
S550 O-Glycosylation
T560 Phosphorylation Q16539 (MAPK14) , P28482 (MAPK1)
K597 Acetylation
T648 O-Glycosylation
T649 Phosphorylation Q13131 (PRKAA1)
S651 Phosphorylation
S654 O-Glycosylation

Research Backgrounds

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'-TT[G/A]TTTTG-3' and the related Daf-16 family binding element (DBE) with consensus sequence 5'-TT[G/A]TTTAC-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. 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. In hepatocytes, promotes gluconeogenesis by acting together with PPARGC1A and CEBPA to activate the expression of genes such as IGFBP1, G6PC and PCK1. 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 (By similarity).

PTMs:

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 (By similarity). 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.

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.

Ubiquitinated by SKP2. Ubiquitination leads to proteasomal degradation.

Methylation inhibits AKT1-mediated phosphorylation at Ser-256 and is increased by oxidative stress.

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.

Subcellular Location:

Cytoplasm. Nucleus.
Note: Shuttles between the cytoplasm and nucleus. Largely nuclear in unstimulated cells. In osteoblasts, colocalizes with ATF4 and RUNX2 in the nucleus (By similarity). 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-proteosomal 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.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Ubiquitous.

Subunit Structure:

Interacts with LRPPRC. Interacts with RUNX2; the interaction inhibits RUNX2 transcriptional activity and mediates the IGF1/insulin-dependent BGLAP expression in osteoblasts Interacts with PPP2R1A; the interaction regulates the dephosphorylation of FOXO1 at Thr-24 and Ser-256 leading to its nuclear import. Interacts (acetylated form) with PPARG. Interacts with XBP1 isoform 2; this interaction is direct and leads to FOXO1 ubiquitination and degradation via the proteasome pathway (By similarity). Interacts with NLK. Interacts with SIRT1; the interaction results in the deacetylation of FOXO1 leading to activation of FOXO1-mediated transcription of genes involved in DNA repair and stress resistance. Binds to CDK1. Interacts with the 14-3-3 proteins, YWHAG and YWHAZ; the interactions require insulin-stimulated phosphorylation on Thr-24, promote nuclear exit and loss of transcriptional activity. Interacts with SKP2; the interaction ubiquitinates FOXO1 leading to its proteosomal degradation. The interaction requires the presence of KRIT1. Interacts (via the C-terminal half) with ATF4 (via its DNA-binding domain); the interaction occurs in osteoblasts, regulates glucose homeostasis via suppression of beta-cell proliferation and subsequent decrease in insulin production. Interacts with PRMT1; the interaction methylates FOXO1, prevents PKB/AKT1 phosphorylation and retains FOXO1 in the nucleus. Interacts with EP300 and CREBBP; the interactions acetylate FOXO1. Interacts with SIRT2; the interaction is disrupted in response to oxidative stress or serum deprivation, leading to increased level of acetylated FOXO1, which promotes stress-induced autophagy by stimulating E1-like activating enzyme ATG7. Interacts (acetylated form) with ATG7; the interaction is increased in response to oxidative stress or serum deprivation and promotes the autophagic process leading to cell death. Interacts (via the Fork-head domain) with CEBPA; the interaction increases when FOXO1 is deacetylated. Interacts with WDFY2. Forms a complex with WDFY2 and AKT1 (By similarity). Interacts with CRY1 (By similarity).

Research Fields

· Cellular Processes > Cell growth and death > Cellular senescence.   (View pathway)

· Environmental Information Processing > Signal transduction > FoxO signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > AMPK signaling pathway.   (View pathway)

· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.

· Human Diseases > Cancers: Specific types > Prostate cancer.   (View pathway)

· Organismal Systems > Aging > Longevity regulating pathway.   (View pathway)

· Organismal Systems > Aging > Longevity regulating pathway - multiple species.   (View pathway)

· Organismal Systems > Endocrine system > Insulin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Glucagon signaling pathway.

References

1). The 15-hydroxyprostaglandin dehydrogenase inhibitor SW033291 ameliorates abnormal hepatic glucose metabolism through PGE2–EP4 receptor–AKT signaling in a type 2 diabetes mellitus mouse model. Cellular Signalling, 2023 (PubMed: 37164143) [IF=4.8]

2). Tea seed saponin‑reduced extract ameliorates palmitic acid‑induced insulin resistance in HepG2 cells. Molecular medicine reports, 2024 (PubMed: 38099345) [IF=3.4]

Application: WB    Species: Human    Sample: HepG2 cells

Figure 6. Effects of TSSRE on phosphorylation of (A) FOXO1 and (B) PEPCK in HepG2 cells. HepG2 cells were treated with normal-(5.5 mM) or high-concentration (30 mM) glucose plus 0.25 mM PA in the absence or presence of TSSRE for 24 h and subsequently treated with insulin (100 nM) for 30 min. *P

3). Glucocorticoids induce osteonecrosis of the femoral head in rats via PI3K/AKT/FOXO1 signaling pathway. PeerJ, 2022 (PubMed: 35529482) [IF=2.7]

Application: WB    Species: Mice    Sample: MC3T3-E1 cells

Figure 4 Dex inhibited the activities of PI3K/AKT signaling pathways in MC3T3-E1 cells. (A–B) Western blotting was performed to analyze the expression levels of PI3K, p-PI3K, AKT, p-AKT, FOXO1, and p-FOXO1 in MC3T3-E1 cells after Dex treatment (200 µM, 48 h). Con: control group. All data in vivo and in vitro experiments are triplicate biological replicates. * P < 0.05, ** P < 0.01, *** P < 0.0001.

Restrictive clause

 

Affinity Biosciences tests all products strictly. Citations are provided as a resource for additional applications that have not been validated by Affinity Biosciences. Please choose the appropriate format for each application and consult Materials and Methods sections for additional details about the use of any product in these publications.

For Research Use Only.
Not for use in diagnostic or therapeutic procedures. Not for resale. Not for distribution without written consent. Affinity Biosciences will not be held responsible for patent infringement or other violations that may occur with the use of our products. Affinity Biosciences, Affinity Biosciences Logo and all other trademarks are the property of Affinity Biosciences LTD.