Purified anti-FOXO1 Antibody

Pricing & Availability
Clone
W20064D (See other available formats)
Regulatory Status
RUO
Other Names
Forkhead Box O1, FOXO1A, FKH1, FKHR
Isotype
Rat IgG2b, κ
Ave. Rating
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Product Citations
publications
W20064D_PURE_FOXO1_Antibody_1_WB_102621.png
Whole cell extracts (15 µg total protein) from HEL, MOLT4 (negative controls), Hela, Daudi (positive controls) and EL4 cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 μg/mL (1:500 dilution) purified anti-FOXO1 antibody (clone W20064D) overnight at 4 °C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker.
  • W20064D_PURE_FOXO1_Antibody_1_WB_102621.png
    Whole cell extracts (15 µg total protein) from HEL, MOLT4 (negative controls), Hela, Daudi (positive controls) and EL4 cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 μg/mL (1:500 dilution) purified anti-FOXO1 antibody (clone W20064D) overnight at 4 °C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker.
  • W20064D_PURE_FOXO1_Antibody_2_ICFC_102621.png
    Daudi cells (filled histogram, positive control) and MOLT-4 cells (open histogram, negative control) were fixed with Fixation Buffer (Cat. No. 420801), permeabilized using True-Phos™ Perm Buffer (Cat. No. 425401), and intracellularly stained with purified anti-FOXO1 antibody (clone W20064D) followed by PE goat anti-rat IgG antibody (Cat. No. 405406).
  • W20064D_PURE_FOXO1_Antibody_3_ICC_102621.png
    ICC staining of purified anti-FOXO1 antibody (clone W20064D) on untreated serum-starved HeLa cells (-) and serum-starved HeLa cells treated with 50 nM insulin for 30 minutes (+) to induce translocation of FOXO1 from the nucleus to the cytoplasm. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with methanol for 10 minutes, and blocked with 5% FBS for 60 minutes. The cells were then incubated with 5 µg/mL of the primary antibody for overnight at 4°C, followed by incubation with 2 µg/mL of Alexa Fluor® 594 goat anti-rat IgG (Cat. No. 405422) for one hour at room temperature. Nuclei were counterstained with DAPI, and the slides were mounted with ProLong™ Gold Antifade Mountant. The image was captured with a 60X objective. Scale bar: 10 µm
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947201 25 µg 80,00€
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947202 100 µg 200,00€
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Description

Forkhead box protein O1 (FOXO1) is a member of the Forkhead box class O family of transcription factors. As such, it is involved in the regulation of a wide range of vital cellular processes including cell cycle progression, differentiation, proliferation, DNA repair, stress response, and apoptosis. FOXO1 is activated via serine and threonine phosphorylation. Phosphorylation is triggered by external stimuli including growth factors, cytokines, and insulin and can be PKB/Akt-mediated as part of the P13K-Akt/SGK pathway on Thr24, Ser256, and Ser319, mediated by CDK2 or CK1 on Ser249 and Ser322/325, respectively, or induced by serum and glucocorticoid-inducible kinase or NFkB kinase. Upon phosphorylation, FOXO1, which is localized to the nucleus, translocates to the cytoplasm following binding to 14-3-3 proteins. In addition to phosphorylation, FOXO1 can be activated via acetylation of its lysine residues by coactivator p300. FOXO1 is vital to the regulation of metabolism through the promotion of glucose homeostasis and gluconeogenic enzyme expression, adipose tissue and skeletal muscle formation, and oxidative stress regulation. FOXO1 may inhibit cell cycle progression and induce apoptosis in cancer cells.

Product Details
Technical Data Sheet (pdf)

Product Details

Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Partial recombinant human FOXO1 protein
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICFC, ICC - Verified

Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.125 - 1.0 µg/mL. For immunocytochemistry, a concentration of 5.0 μg/mL is recommended. For flow cytometric staining, the suggested use of this reagent is ≤ 0.125 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This clone was tested for ICC using serum-starved HeLa cells treated with insulin and untreated serum-starved HeLa cells. Staining was expected to localize to the nucleus prior to treatment and localize to the cytoplasm following treatment. Three fix/perm methods were used: methanol fixation or 4% PFA fixation followed by permeabilization with either methanol or Triton X-100.  Only PFA fixation followed by methanol permeabilization produced a strong signal and the expected shift in localization.

This clone was tested for IHC on human melanoma tissue using Biolegend’s DAB Chromagen. No staining was produced.

Product Citations

Antigen Details

Structure
FOXO1 is a 655 amino acid protein with a predicted molecular weight of 69 kD.
Distribution

Ubiquitously expressed /Nucleus and Cytoplasm

Function
Transcription Factor
Interaction
RET signaling: PDGF, EGFR, DAP12, MAPK, NGF
Cytokine Receptor Gamma-Chain Family signaling: IL-21, IL-7, IL-9, IL-15, IL-2, IL-4, adipokines, insulin, thrombopoietin
GAB1 signalosome: PI3K/AKT, GAB1
Immune response IL-23 signaling: Tie2
Cell Type
B cells, Tregs
Biology Area
Cell Biology, Cell Cycle/DNA Replication, Immunology, Transcription Factors
Molecular Family
Nuclear Markers
Antigen References
  1. Greer E and Brunet A. 2005. Oncogene. 24:7410.
  2. Gross D, et al. 2008. Oncogene. 27:2320.
  3. Perrot V and Matthew R. 2005. Mol. Endocrinol. 19:2283.
  4. Tzivion G, et al. 2011. BBA. 1813:1938.
  5. Yang Y, et al. 2005. EMBO. 24:1021.
Gene ID
2308 View all products for this Gene ID
UniProt
View information about FOXO1 on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 10.26.2021

For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

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