Key features and details
- Rabbit polyclonal to FOXO3A
- Suitable for: IHC-P, WB, ICC/IF
- Reacts with: Mouse, Human
- Isotype: IgG
Product nameAnti-FOXO3A antibody
See all FOXO3A primary antibodies
DescriptionRabbit polyclonal to FOXO3A
Tested applicationsSuitable for: IHC-P, WB, ICC/IFmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat
General notesStorage in frost-free freezers is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilutions should be discarded if not used within 12 hours.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.097% Sodium azide
Constituents: 0.0268% PBS, 1% BSA
Concentration information loading...
PurityImmunogen affinity purified
ChIP Related Products
Our Abpromise guarantee covers the use of ab12162 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 10 µg/ml.|
|WB||1/2500. Detects a band of approximately 70-75 kDa (predicted molecular weight: 71 kDa). In some preparations there is an additional band at 35 kDa.
A customer reported the antibody did not detect endogenous FOX03A in human cancer cells (see enquiries).
FunctionTranscriptional 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'.
Involvement in diseaseNote=A chromosomal aberration involving FOXO3 is found in secondary acute leukemias. Translocation t(6;11)(q21;q23) with MLL/HRX.
Sequence similaritiesContains 1 fork-head DNA-binding domain.
modificationsIn 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 on Ser-209 upon oxidative stress, which leads to dissociation from YWHAB/14-3-3-beta and nuclear translocation. Phosphorylated by PIM1.
Cellular localizationCytoplasm > cytosol. Nucleus. Translocates to the nucleus upon oxidative stress and in the absence of survival factors.
- Information by UniProt
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue labelling FOXO3a with ab12162 at 10 µg/mL.
Immunocytochemistry/immunofluorescence analysis of human cervix carcinoma cells labelling FOXO3A with ab[12162 at 1/200 dilution.
ab12162 staining FOXO3A in Rat fibroblast cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.03% Triton X-100 and blocked with 5% serum for 10 minutes at 25°C. Samples were incubated with primary antibody (1/300) for 1 hour at 25°C.
Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human colon carcinoma tissue, staining FOXO3A with ab12162.
Tissue was fixed with formaldehyde and blocked with 1% serum for 1 hour at 24°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100 in diluent) for 12 hours at 4°C. A biotinylated goat anti-rabbit polyclonal IgG (1/200) was used as the secondary antibody.
Anti-FOXO3A antibody (ab12162) at 1/1500 dilution + Mouse muscle nuclear cell lysate at 40 µg
Donkey anti-rabbit HRP conjugate (undiluted)
Developed using the ECL technique.
Predicted band size: 71 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 1 minute
For protein extraction, murine BaF3Flt-3,Il-7 cells were washed in ice-cold PBS and immediately resuspended in lysis buffer containing 25 mm Tris (pH 7.5), 1% Triton X-100, 150 mm NaCl, 1 mm DTT, 1 mm EDTA (pH 8.0), 1× protease inhibitor mixture and 1 mm Na3VO4. Samples were centrifuged 1000 × g for 2 minutes and supernatant was mixed with 1× LDS sample buffer and 0.01 m DTT. Samples were heated at 70 °C for 10 minutes before separated by SDS-PAGE and blotted onto a PVDF transfer membrane. Membranes were blocked in 5% milk PBS/Tween or 5% BSA TBS/Tween for 1 hour at room temperature and next incubated with primary antibodies for 1 hour at room temperature or at 8 °C overnight. Following block, membranes were incubated with HRP-conjugated secondary antibody for 1 hour at room temperature. Blots were developed by enhanced chemiluminescence.
ab12162 has been referenced in 41 publications.
- Wu Y et al. Sepiapterin reductase promotes hepatocellular carcinoma progression via FoxO3a/Bim signaling in a nonenzymatic manner. Cell Death Dis 11:248 (2020). PubMed: 32312975
- Alasiri G et al. Regulation of PERK expression by FOXO3: a vulnerability of drug-resistant cancer cells. Oncogene 38:6382-6398 (2019). PubMed: 31312024
- Lim SW et al. Ginseng increases Klotho expression by FoxO3-mediated manganese superoxide dismutase in a mouse model of tacrolimus-induced renal injury. Aging (Albany NY) 11:5548-5569 (2019). PubMed: 31400753
- Cao L et al. Genotoxic stress-triggered ß-catenin/JDP2/PRMT5 complex facilitates reestablishing glutathione homeostasis. Nat Commun 10:3761 (2019). PubMed: 31434880
- Li J et al. Knockdown of FOXO3a induces epithelial-mesenchymal transition and promotes metastasis of pancreatic ductal adenocarcinoma by activation of the ß-catenin/TCF4 pathway through SPRY2. J Exp Clin Cancer Res 38:38 (2019). PubMed: 30691517