Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) Recombinant Rabbit Monoclonal Antibody [PSH06-00]

Overview

Product Name

Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) Recombinant Rabbit Monoclonal Antibody [PSH06-00] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell, FC

Positive Control

Jurkat treated with 100nM Calyculin A for 40 minutes cell lysate, NIH/3T3 starved for 24 hours then add 100nM Calyculin A for 30 minutes cell lysate, Jurkat cells treated with 100nM Calyculin A for 40 minutes.

Conjugation

unconjugated

Clone Number

PSH06-00

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

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

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

WB
1:1,000

IF-Cell
1:1,000

FC
1:1,000

Target

Function

FOX (forkhead box) proteins are a family of transcription factors that play important roles in regulating the expression of genes involved in cell growth, proliferation, differentiation, and longevity. Many FOX proteins are important to embryonic development. FOX proteins also have pioneering transcription activity by being able to bind condensed chromatin during cell differentiation processes. The defining feature of FOX proteins is the forkhead box, a sequence of 80 to 100 amino acids forming a motif that binds to DNA. This forkhead motif is also known as the winged helix, due to the butterfly-like appearance of the loops in the protein structure of the domain. Forkhead proteins are a subgroup of the helix-turn-helix class of proteins. Some FOX genes are downstream targets of the hedgehog signaling pathway, which plays a role in the development of basal cell carcinomas. Members of class O (FOXO- proteins) regulate metabolism, cellular proliferation, stress tolerance and possibly lifespan. The activity of FoxO is controlled by post-translational modifications, including phosphorylation, acetylation and ubiquitination.

Background References

1. Adiguzel D et al. FoxO1 is a cell-specific core transcription factor for endometrial remodeling and homeostasis during menstrual cycle and early pregnancy. Hum Reprod Update. 2021 Apr

2. Hao Y et al. p300 arrests intervertebral disc degeneration by regulating the FOXO3/Sirt1/Wnt/beta-catenin axis. Aging Cell. 2022 Aug

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: Q12778 Human

SwissProt: O43524 Human

SwissProt: P98177 Human

SwissProt: A8MYZ6 Human

SwissProt: Q9R1E0 Mouse

SwissProt: Q9WVH4 Mouse

SwissProt: Q9WVH3 Mouse

SwissProt: Q70KY4 Mouse

Synonyms

FKH 1 antibody

FKH1 antibody

FKHR antibody

Forkhead (Drosophila) homolog 1 (rhabdomyosarcoma) antibody

Forkhead box O1 antibody

Forkhead box protein O1 antibody

Forkhead box protein O1A antibody

Forkhead in rhabdomyosarcoma antibody

Forkhead, Drosophila, homolog of, in rhabdomyosarcoma antibody

FoxO transcription factor antibody

Expand

Images

☑ Cell treatment (CT)

Western blot analysis of Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) on different lysates with Rabbit anti-Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) antibody (HA751040) at 1/1,000 dilution.

Lane 1: Jurkat cell lysate
Lane 2: Jurkat treated with 20μM LY294002 for 2 hours cell lysate
Lane 3: Jurkat treated with 100nM Calyculin A for 40 minutes cell lysate
Lane 4: NIH/3T3 starved for 24 hours cell lysate
Lane 5: NIH/3T3 starved for 24 hours then add 100nM Calyculin A for 30 minutes cell lysate

Lysates/proteins at 20 µg/Lane.

Observed band size: 55-150 kDa

Exposure time: 20 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751040) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Cell treatment (CT)

Flow cytometric analysis of Jurkat cells treated with 20μM LY294002 for 2 hours (left, negative) and Jurkat cells treated with 100nM Calyculin A for 40 minutes (right, positive) labeling Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26).

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751040, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
☑ Cell treatment (CT)

Immunocytochemistry analysis of Jurkat cells treated with 20μM LY294002 for 2 hours (upper, negative) and Jurkat cells treated with 100nM Calyculin A for 40 minutes (lower, positive) labeling Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) with Rabbit anti-Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) antibody (HA751040) at 1/1,000 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) antibody (HA751040) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"