PRODUCT CODE: ET1701-67

FOXO4 Recombinant Rabbit Monoclonal Antibody [JJ09-11] (ET1701-67)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

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ICC staining of FOXO4 in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-67, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of FOXO4 in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-67, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of FOXO4 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-67, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of FOXO4 in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-67, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-FOXO4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-67, 1/50)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of FOXO4 was done on 293 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1701-67, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ICC staining of FOXO4 in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-67, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Applications

  • WB

  • ICC

  • IF

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

FOXO4 Recombinant Rabbit Monoclonal Antibody [JJ09-11] (ET1701-67)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

293T, Hela, HepG2, human placenta tissue, 293.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JJ09-11

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

54 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • FC

  • 1:50-1:100

  • ICC/IF

  • 1:100-1:500

TARGET

UNIPROT #

PROTEIN NAME

FOXO4

SYNONYMS

AFX antibody; AFX1 antibody; Afxh antibody; ALL1-fused gene from X chromosome antibody; Fork head domain transcription factor AFX1 antibody; Forkhead box O4 antibody; Forkhead box protein O4 antibody; FOXO 4 antibody; Foxo4 antibody; FOXO4_HUMAN antibody; MGC117660 antibody; MGC120490 antibody; Mixed lineage leukemia, translocated to, 7 antibody; MLLT7 antibody; Myeloid/lymphoid or mixed lineage leukemia (trithorax homolog, Drosophila); translocated to, 7 antibody; Myeloid/lymphoid or mixed lineage leukemia, translocated to, 7 antibody; RGD1561201 antibody

TISSUE SPECIFICITY

Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. Isoform zeta is most abundant in the liver, kidney, and pancreas.

POST-TRANSLATIONAL MODIFICATION

Acetylation by CREBBP/CBP, which is induced by peroxidase stress, inhibits transcriptional activity. Deacetylation by SIRT1 is NAD-dependent and stimulates transcriptional activity.; Phosphorylation by PKB/AKT1 inhibits transcriptional activity and is responsible for cytoplasmic localization. May be phosphorylated at multiple sites by NLK.; Monoubiquitinated; monoubiquitination is induced by oxidative stress and reduced by deacetylase inhibitors; results in its relocalization to the nucleus and its increased transcriptional activity. Deubiquitinated by USP7; deubiquitination is induced by oxidative stress; enhances its interaction with USP7 and consequently, deubiquitination; increases its translocation to the cytoplasm and inhibits its transcriptional activity. Hydrogene-peroxide-induced ubiquitination and USP7-mediated deubiquitination have no major effect on its protein stability.

SUBCELLULAR LOCATION

Nucleus, Cytoplasm.

FUNCTION

FKHR (for forkhead in rhabdomyosarcoma), FKHRL1, and AFX1 are members of a subfamily of the forkhead family of transcription factors. AFX1, also known as FoxO4, is expressed in a wide variety of tissues and, like other FKHR proteins, AFX1 contains a single forkhead domain and serine-proline-rich region, which mediate DNA binding. AFX1-mediated transcriptional activation is regulated by the serine/threonine kinase Akt1, which phosphorylates AFX1 and in turn, sequesters AFX1 in the cytosol, thereby blocking nuclear localization and DNA binding. Genetic mutations in FKHR genes, including the t(2;13) and t(1;3) translocations, are commonly found in alveolar rhabdomyosarcomas. Additionally, the t(x;11) translocation of the AFX1 gene, which involves the fusion of a serine-proline-rich sequence of AFX1 to the carboxy terminus of a truncated MLL, results in acute lymphocytic leukemia.