Key features and details
- Rabbit polyclonal to FOXO3A - ChIP Grade
- Suitable for: IHC-P, IP, IHC-FoFr, WB, ICC/IF, ChIP
- Reacts with: Mouse, Human, Pig
- Isotype: IgG
Product nameAnti-FOXO3A antibody - ChIP Grade
See all FOXO3A primary antibodies
DescriptionRabbit polyclonal to FOXO3A - ChIP Grade
Tested applicationsSuitable for: IHC-P, IP, IHC-FoFr, WB, ICC/IF, ChIPmore details
Species reactivityReacts with: Mouse, Human, Pig
Predicted to work with: Rat
- Recombinant Human FOXO3A protein (ab114191) can be used as a positive control in WB. FOXO3A transfected 293T cells
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 at an assay dependent concentration.|
|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).
|ICC/IF||Use at an assay dependent concentration.|
|ChIP||Use at an assay dependent concentration.|
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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Pig coronary artery endothelial cells were cross-linked for 10 minutes at room temperature by adding 37% formaldehyde to the culture medium. The fixed cells were lysed and the chromatin was isolated. For ChIP, sheared chromatin was incubated with Anti-FOXO3A antibody - ChIP Grade (ab12162). The cells were incubated with wortmannin (30 nM), SB 203580 (10 µM) and SP 600125 (30 µM) for 30 minutes before the addition of CGJ for 30 minutes. n = 4 different experiments. *P<0.05 versus control, #P<0.05 versus CGJ treatment. Endothelial cells were exposed to CGJ for 30 minutes and then chromatin-bound DNA was immunoprecipitated with an antibody against FoxO3a.
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 - ChIP Grade (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 staining FOXO3A in human brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with formaldehyde, permeablized with 0.01% Triton-X 100 and blocked with 5% serum for 4 hours at 25°C. The sample was incubated with primary antibody (1/300) at 4°C for 10 hours. An Alexa Fluor® 594-conjugated donkey anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.
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