Rabbit Anti-Phospho-FoxO3a (Ser318 + Ser321) antibody

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AF6q21; AF6q21 protein; DKFZp781A0677; FKHR2; FKHRL 1; FKHRL1; FKHRL1P2; Forkhead (Drosophila) homolog (rhabdomyosarcoma) like 1; Forkhead box O3; Forkhead box O3A; Forkhead box protein O3; Forkhead box protein O3A; Forkhead Drosophila homolog of in rhabd

Cat:

SL3138R

Species Reactivity：

Human,Rat,(predicted: Mouse,Chicken,Dog,Pig,Cow,Horse,Sheep,)

Immunogen：

KLH conjugated Synthesised phosphopeptide derived from human FoxO3a around the phosphorylation site of Ser318/321:NA(p-S)TV(p-S)GR

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Polyclonal

Isotype：

IgG

Applications：

WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500Flow-Cyt=1ug/TestIF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Rabbit

Product Overview：

Sample: MCF-7(Human) Cell Lysate at 30 ugPrimary: Anti-Phospho-FoxO3a (Ser318 + Ser321) (SL3138R) at 1/500 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 71 kDObserved band size: 71 kD Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by microwave in sodium citrate buffer (pH6.0) ; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (3% BSA) at RT for 30min; Antibody incubation with (Phospho-FoxO3a (Ser318 + Ser321)) Polyclonal Antibody, Unconjugated (SL3138R) at 1:400 overnight at 4℃, followed by conjugation to the secondary antibody (labeled with HRP)and DAB staining.Blank control:THP-1. Primary Antibody (green line): Rabbit Anti-Phospho-FoxO3a (Ser318 + Ser321) antibody (SL3138R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-FITCDilution: 0.5μg /test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.phosphopeptidenon phosphopeptide

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Unit:

Price: $256.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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This gene belongs to the forkhead family of transcription factors which are characterized by a distinct forkhead domain. This gene likely functions as a trigger for apoptosis through expression of genes necessary for cell death. Translocation of this gene with the MLL gene is associated with secondary acute leukemia. Alternatively spliced transcript variants encoding the same protein have been observed. [provided by RefSeq, Jul 2008]

Function:
Transcriptional activator which triggers apoptosis in the absence of survival factors, including neuronal cell death upon oxidative stress. Recognizes and binds to the DNA sequence 5'-[AG]TAAA[TC]A-3'. 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.

Subunit:
Interacts with YWHAB/14-3-3-beta and YWHAZ/14-3-3-zeta, which are required for cytosolic sequestration. Upon oxidative stress, interacts with STK4/MST1, which disrupts interaction with YWHAB/14-3-3-beta and leads to nuclear translocation. Interacts with PIM1.

Subcellular Location:
Cytoplasm, cytosol. Nucleus. Note=Translocates to the nucleus upon oxidative stress and in the absence of survival factors.

Tissue Specificity:
Ubiquitous.

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. 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.

DISEASE:
Note=A chromosomal aberration involving FOXO3 is found in secondary acute leukemias. Translocation t(6;11)(q21;q23) with MLL/HRX.

Similarity:
Contains 1 fork-head DNA-binding domain.

SWISS:
O43524

Gene ID:
2309

Database links:

Entrez Gene: 2309 Human

Entrez Gene: 56484 Mouse

Entrez Gene: 294515 Rat

Omim: 602681 Human

SwissProt: O43524 Human

SwissProt: Q9WVH4 Mouse

Unigene: 220950 Human

Unigene: 338613 Mouse

Unigene: 24593 Rat

FOXO3a是FOX(forkhead box)蛋白家族的一个重要成员,与细胞转化、肿瘤的发生发展及其血管生成有关,是一种已知的控制细胞循环和细胞死亡的蛋白质。 Picture

Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by microwave in sodium citrate buffer (pH6.0) ; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (3% BSA) at RT for 30min; Antibody incubation with (Phospho-FoxO3a (Ser318 + Ser321)) Polyclonal Antibody, Unconjugated (SL3138R) at 1:400 overnight at 4℃, followed by conjugation to the secondary antibody (labeled with HRP)and DAB staining.

Blank control:THP-1.
Primary Antibody (green line): Rabbit Anti-Phospho-FoxO3a (Ser318 + Ser321) antibody (SL3138R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 0.5μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

phosphopeptide non phosphopeptide

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