Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) Recombinant Rabbit Monoclonal Antibody [PSH06-00]
cat.: HA722499
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH06-00 |
| Form: | Liquid |
| Storage condition: | 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. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Isotype: | IgG |
| 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. |
| Subcellular location: | Cytoplasm, Nucleus. |
| Recommended Dilutions:
WB IF-Cell FC |
1:1,000 1:1,000 1:1,000 |
| Uniprot #: | SwissProt: Q12778 Human | O43524 Human | P98177 Human | A8MYZ6 Human | Q9R1E0 Mouse | Q9WVH4 Mouse | Q9WVH3 Mouse | Q70KY4 Mouse |
| Alternative names: | FKH 1 FKH1 FKHR Forkhead (Drosophila) homolog 1 (rhabdomyosarcoma) Forkhead box O1 Forkhead box protein O1 Forkhead box protein O1A Forkhead in rhabdomyosarcoma Forkhead, Drosophila, homolog of, in rhabdomyosarcoma FoxO transcription factor foxo1 FOXO1_HUMAN FOXO1A OTTHUMP00000018301 AF6q21 AF6q21 protein DKFZp781A0677 FKHR2 FKHRL 1 FKHRL1 FKHRL1P2 Forkhead (Drosophila) homolog (rhabdomyosarcoma) like 1 Forkhead box O3 Forkhead box O3A Forkhead box protein O3 Forkhead box protein O3A Forkhead Drosophila homolog of in rhabdomyosarcoma like 1 Forkhead homolog (rhabdomyosarcoma) like 1 Forkhead in rhabdomyosarcoma like 1 Forkhead in rhabdomyosarcoma-like 1 FOX O3A FOXO2 foxo3 FOXO3_HUMAN FOXO3A MGC12739 MGC31925 AFX AFX1 Afxh ALL1-fused gene from X chromosome Fork head domain transcription factor AFX1 Forkhead box O4 Forkhead box protein O4 FOXO 4 Foxo4 FOXO4_HUMAN MGC117660 MGC120490 Mixed l...... |
Images
|
Fig1:
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 (HA722499) 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 (HA722499) 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. |
|
Fig2:
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 (HA722499, 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). |
|
Fig3:
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 (HA722499) 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 (HA722499) 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.