FOXO3A Monoclonal antibody

• Featured Product
• KD/KO Validated

FOXO3A Monoclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Host / Isotype

Mouse / IgG2a

Reactivity

human, mouse, rat and More (2)

Applications

WB, IHC, IF/ICC, IP, CoIP, ChIP, ELISA

Conjugate

Unconjugated

Publications

46

CloneNo.

1F12D11

Cat no : 66428-1-Ig

Synonyms

AF6q21, AF6q21 protein, FKHRL1, FKHRL1P2, forkhead box O3, Forkhead box protein O3, FOXO2, FOXO3, FOXO3A

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Validation Data Gallery

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  WB analysis using 66428-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66428-1-Ig (FOXO3A antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Beta Actin Monoclonal antibody (HRP-66009) as loading control.

• 

  WB analysis using 66428-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66428-1-Ig (FOXO3A antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Beta Actin Monoclonal antibody (HRP-66009) as loading control.

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  WB analysis of HEK-293 using 66428-1-Ig

  WB result of FOXO3A antibody (66428-1-Ig, 1:12,000) with si-Control and si-FOXO3A transfected HEK293 cells.

• 

  WB analysis using 66428-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66428-1-Ig (FOXO3A antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  IP experiment of HEK-293 using 66428-1-Ig

  IP result of anti-FOXO3A (IP:66428-1-Ig, 4ug; Detection:66428-1-Ig 1:1000) with HEK-293 cells lysate 2200 ug.

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  IHC staining of human breast cancer using 66428-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66428-1-Ig (FOXO3A antibody) at dilution of 1:500 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 66428-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66428-1-Ig (FOXO3A antibody) at dilution of 1:500 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 66428-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66428-1-Ig (FOXO3A antibody) at dilution of 1:500 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human stomach cancer using 66428-1-Ig

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 66428-1-Ig (FOXO3A Antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human stomach cancer using 66428-1-Ig

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 66428-1-Ig (FOXO3A Antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human ovary tumor using 66428-1-Ig

  Immunohistochemical analysis of paraffin-embedded human ovary tumor tissue slide using 66428-1-Ig (FOXO3A antibody) at dilution of 1:500 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human ovary tumor using 66428-1-Ig

  Immunohistochemical analysis of paraffin-embedded human ovary tumor tissue slide using 66428-1-Ig (FOXO3A antibody) at dilution of 1:500 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 66428-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66428-1-Ig (FOXO3A antibody) at dilution of 1:500 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HepG2 using 66428-1-Ig

  Immunofluorescent analysis of (4% PFA ) fixed HepG2 cells using 66428-1-Ig(FOXO3A antibody) at dilution of 1:50 and Alexa Fluor 594-conjugated Goat Anti-mouse IgG(H+L).

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Tested Applications

• Positive WB detected in: HEK-293 cells, PC-12 cells, MCF-7 cells, HepG2 cells, Jurkat cells, K-562 cells, HSC-T6 cells, NIH/3T3 cells, Neuro-2a cells
• Positive IP detected in: HEK-293 cells
• Positive IHC detected in: human breast cancer tissue, human lung cancer tissue, human ovary tumor tissue, human stomach cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:250-1:1000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 7 publications below
• WB: See 39 publications below
• IHC: See 7 publications below
• IF: See 13 publications below
• IP: See 5 publications below
• CoIP: See 5 publications below
• ChIP: See 2 publications below

Product Information

66428-1-Ig targets FOXO3A in WB, IHC, IF/ICC, IP, CoIP, ChIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, zebrafish, macaque
• Host / Isotype: Mouse / IgG2a
• Class: Monoclonal
• Type: Antibody
• Immunogen: FOXO3A fusion protein Ag1289
• Full Name: forkhead box O3
• Calculated Molecular Weight: 673 aa, 71 kDa
• Observed Molecular Weight: 85-90 kDa
• GenBank Accession Number: BC021224
• Gene Symbol: FOXO3A
• Gene ID (NCBI): 2309
• RRID: AB_2881799
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

In mammalian cells, FoxO factors act as potent transcriptional activators that upregulate the expression of programs of genes involved in stress resistance, cell cycle arrest, differentiation, apoptosis, autophagy, and metabolism. FOXO3 has a key role in the maintenance of adult neural and hematopoietic stem cells. It also associates with extreme longevity in humans and regulates the homeostasis of adult stem cell pools in mammals, which may contribute to longevity. Phosphorylation inhibits FOXO3 nuclear translocation that is essential for its regulation and function. The calculated molecular weight of FOXO3 is 71 kDa,but some experiments analysis using a FOXO3a specific antibody showed the expression of a 90 kDa fulllength FOXO3a protein (Full) and a 40 kDa processed FOXO3a product (Short). 90 kDa FOXO3a protein is a phosphorylated via activation of PI3K (phosphatidylinositol 3-kinase) and Akt after INS stimulation, and translocate from the nucleus to the cytoplasm. (PMID: 19427386)

Publications

Species	Application	Title
human	WB,IHC,IP	Autophagy; CDK9 inhibition blocks the initiation of PINK1-PRKN-mediated mitophagy by regulating the SIRT1-FOXO3-BNIP3 axis and enhances the therapeutic effects involving mitochondrial dysfunction in hepatocellular carcinoma.; Authors - Jingyue Yao
human	WB	Clin Immunol; METTL14-m6A-FOXO3a axis regulates autophagy and inflammation in ankylosing spondylitis; Authors - Yuting Chen ; KD Validated
human	WB	J Control Release; Next generation miRNA inhibition using short anti-seed PNAs encapsulated in PLGA nanoparticles.; Authors - Shipra Malik
human	WB	Transl Psychiatry; Psychosocial and behavioral problems of children and adolescents in the early stage of reopening schools after the COVID-19 pandemic: a national cross-sectional study in China.; Authors - Lin Wang
human	ChIP	J Dent Res; FOXO3 Mediates Tooth Movement by Regulating Force-Induced Osteogenesis.; Authors - A Jin
human	WB	Int J Biol Sci; ATIC inhibits autophagy in hepatocellular cancer through the AKT/FOXO3 pathway and serves as a prognostic signature for modeling patient survival.; Authors - Hao Zhang