Human/Mouse FoxO3 Antibody

  
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human and mouse FoxO3 in Western blots.
  • Source
    Monoclonal Mouse IgG2A Clone # 648716
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human FoxO3
    Ala373-Gly673
    Accession # O43524
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Immunocytochemistry
    5-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human and Mouse FoxO3 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, MCF‑7 human breast cancer cell line, Jurkat human acute T cell leukemia cell line, and NIH‑3T3 mouse embryonic fibroblast cell line. PVDF Membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse FoxO3 Monoclonal Antibody (Catalog # MAB6165) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for FoxO3 at approximately 85 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
FoxO3 in MCF‑7 Human Cell Line. FoxO3 was detected in immersion fixed MCF‑7 human breast cancer cell line using Mouse Anti-Human/Mouse FoxO3 Monoclonal Antibody (Catalog # MAB6165) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: FoxO3
Forkhead box O3 (FoxO3) is a ubiquitously expressed transcriptional regulator that is involved in cellular differentiation, angiogenesis, tumor progression, apoptosis, and the responses to oxidative stress and DNA damage. Phosphorylation of FoxO3 by Akt induces its association with 14-3-3 proteins and its retention in the cytoplasm. In response to the loss of survival factors, dephosphorylation of FoxO3 induces its translocation to the nucleus where it promotes apoptosis. Its level of acetylation is regulated in response to cellular metabolic requirements. Within amino acids 372‑673 (C-terminal to the DNA binding domain), human FoxO3 shares 95% aa sequence identity with mouse and rat FoxO3.
  • Long Name:
    Forkhead Box O3
  • Entrez Gene IDs:
    2309 (Human); 56484 (Mouse); 294515 (Rat)
  • Alternate Names:
    AF6q21 protein; AF6q21; DKFZp781A0677; FKHRL1; FKHRL1MGC31925; FKHRL1P2; forkhead box O3; forkhead box protein O3; forkhead homolog (rhabdomyosarcoma) like 1; Forkhead in rhabdomyosarcoma-like 1; forkhead, Drosophila, homolog of, in rhabdomyosarcoma-like 1; FOXO2; FoxO3; FOXO3A; FOXO3Aforkhead box O3A; MGC12739
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