WB
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 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; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
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 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 at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
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 (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Modification | p-T26 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFWB |
| WB:1:1000 ICC:1:1000 FC:1:1000 |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | FOXO1FOXO3FOXO4FOXO6 |
|---|---|
| Alternative Names | 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 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 foxo1 antibody FOXO1_HUMAN antibody FOXO1A antibody OTTHUMP00000018301 antibody AF6q21 antibody AF6q21 protein antibody DKFZp781A0677 antibody FKHR2 antibody FKHRL 1 antibody FKHRL1 antibody FKHRL1P2 antibody Forkhead (Drosophila) homolog (rhabdomyosarcoma) like 1 antibody Forkhead box O3 antibody Forkhead box O3A antibody Forkhead box protein O3 antibody Forkhead box protein O3A antibody Forkhead Drosophila homolog of in rhabdomyosarcoma like 1 antibody Forkhead homolog (rhabdomyosarcoma) like 1 antibody Forkhead in rhabdomyosarcoma like 1 antibody Forkhead in rhabdomyosarcoma-like 1 antibody FOX O3A antibody FOXO2 antibody foxo3 antibody FOXO3_HUMAN antibody FOXO3A antibody MGC12739 antibody MGC31925 antibody 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 FOXO6 Forkhead box protein O6 antibody |
| Molecular Weight(MW) | 55-150 kDa |
| 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. |
| Cellular Localization | Cytoplasm, Nucleus. |
WB
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 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; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
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 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 at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
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 (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:1000 ICC:1:1000 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
