AF6165

antibody from R&D Systems

Targeting: FOXO3 AF6q21, FKHRL1, FOXO2, FOXO3A

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

AF6165

Provider

R&D Systems

Product name

Human/Mouse FoxO3 Antibody

Antibody type

Polyclonal

Description

Antigen Affinity-purified. Detects human and mouse FoxO3 in Western blots.

Reactivity

Human, Mouse

Host

Sheep

Conjugate

Unconjugated

Antigen sequence

O43524


Isotype

IgG

Vial size

100 ug

Concentration

LYOPH

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.

Western blot

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

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.

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

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.

Immunocytochemistry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

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.