Recombinant Anti-FOXO3A antibody [EPR1950] (ab109629)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1950] to FOXO3A
- Suitable for: Flow Cyt (Intra), WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-FOXO3A antibody [EPR1950]
See all FOXO3A primary antibodies -
Description
Rabbit monoclonal [EPR1950] to FOXO3A -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WBmore details
Unsuitable for: IHC-P or IP -
Species reactivity
Reacts with: Human
Predicted to work with: Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- HEK-293, HEK-293T, MCF7 and SH-SY5Y cell lysates, HeLa cells
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1950 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109629 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB |
1/1000 - 1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 71 kDa).
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
1/1000 - 1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 71 kDa). |
Target
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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'. -
Tissue specificity
Ubiquitous. -
Involvement in disease
Note=A chromosomal aberration involving FOXO3 is found in secondary acute leukemias. Translocation t(6;11)(q21;q23) with MLL/HRX. -
Sequence similarities
Contains 1 fork-head DNA-binding domain. -
Post-translational
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 localization
Cytoplasm > cytosol. Nucleus. Translocates to the nucleus upon oxidative stress and in the absence of survival factors. - Information by UniProt
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Database links
- Entrez Gene: 2309 Human
- Entrez Gene: 294515 Rat
- Omim: 602681 Human
- SwissProt: O43524 Human
- Unigene: 220950 Human
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Alternative names
- AF6q21 antibody
- AF6q21 protein antibody
- DKFZp781A0677 antibody
see all
Images
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All lanes : Anti-FOXO3A antibody [EPR1950] (ab109629) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 :Human FOXO3 (FOXO3A) knockout HEK-293T cell lysate (ab256922)
Lane 3 : Wild-type HEK-293 cell lysate
Lane 4 :Human FOXO3 (FOXO3A) knockout HEK-293 cell lysate (ab261649)
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 82 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab109629 observed at 82 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab109629 was shown to react with FOXO3A in wild-type cells in Western blot with loss of signal observed in FOXO3 knockout cell lines. Wild-type and FOXO3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab109629 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling FOXO3A with unpurified ab109629 at 1/150 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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All lanes : Anti-FOXO3A antibody [EPR1950] (ab109629) at 1/1000 dilution
Lane 1 : Wild-type HEK 293 whole cell lysate
Lane 2 : FOXO3 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 71 kDaLanes 1 - 4: Merged signal (red and green). Green - ab109629 observed at 71 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109629 was shown to recognize FOXO3A in wild-type HEK 293 cells as signal was lost at the expected MW in FOXO3 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and FOXO3 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab109629 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-FOXO3A antibody [EPR1950] (ab109629) at 1/1000 dilution
Lane 1 : MCF7 cell lysate
Lane 2 : SH-SY5Y cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 71 kDa
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (26)
ab109629 has been referenced in 26 publications.
- Zheng Y et al. EOAI, a ubiquitin-specific peptidase 5 inhibitor, prevents non-small cell lung cancer progression by inducing DNA damage. BMC Cancer 23:28 (2023). PubMed: 36611139
- Sun X & Liu B Donepezil ameliorates oxygen-glucose deprivation/reoxygenation-induced brain microvascular endothelial cell dysfunction via the SIRT1/FOXO3a/NF-κB pathways. Bioengineered 13:7760-7770 (2022). PubMed: 35286233
- Sun R & Zheng Q AKT/foxo3a signal pathway mediates the protective mechanism of resveratrol on renal interstitial fibrosis and oxidative stress in rats with unilateral ureteral obstruction. Am J Transl Res 14:1788-1795 (2022). PubMed: 35422955
- Yao F & Zhu Q α-Cyperone Protects Cardiomyocytes against Oxygen-Glucose Deprivation-Induced Inflammation and Oxidative Stress by Akt/FOXO3a/NF-κB Pathway. Dis Markers 2022:8205707 (2022). PubMed: 36072899
- Li H et al. Tubular β-catenin alleviates mitochondrial dysfunction and cell death in acute kidney injury. Cell Death Dis 13:1061 (2022). PubMed: 36539406