Human/Mouse FoxO3 Antibody

Catalog #: AF6165 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF6165
AF6165-SP

Detection of Human FoxO3 by Western Blot.

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Human/Mouse FoxO3 Antibody Summary

Species Reactivity

Human, Mouse

Specificity

Detects human and mouse FoxO3 in Western blots.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human FoxO3
Met372-Gly673
Accession # O43524

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Simple Western

10 µg/mL

MCF‑7 human breast cancer cell line

Immunocytochemistry

5-15 µg/mL

See below

Knockout Validated

FoxO3 is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in FoxO3 knockout HEK293T cell line.

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.

Scientific Data

Western Blot

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Detection of Human FoxO3 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line, MCF-7 human breast cancer cell line, and PC-3 human prostate cancer cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human FoxO3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6165) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for FoxO3 at approximately 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

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FoxO3 in MCF‑7 Human Cell Line. FoxO3 was detected in immersion fixed MCF-7 human breast cancer cell line using Sheep Anti-Human FoxO3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6165) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red, upper panel; Catalog # NL010) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Simple Western

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Detection of Human FoxO3 by Simple Western^TM. Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for FoxO3 at approximately 97 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human/Mouse FoxO3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6165) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Knockout Validated

Western Blot Shows Human FoxO3 Antibody Specificity by Using Knockout Cell Line.

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Western Blot Shows Human FoxO3 Specificity by Using Knockout Cell Line. Western blot shows lysates of HEK293T human embryonic kidney parental cell line and FoxO3 knockout HEK293T cell line (KO). PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse FoxO3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6165) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for FoxO3 at approximately 90 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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 72 kDa 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