Immunofluorescence analysis of FOXO3 was done on 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with FOXO3 Rabbit Polyclonal antibody (PA1805) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing Nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues L N(654) V G N F T G A K Q A S S Q S W V(670) of human FOXO3.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Western Blot (WB)||1:500-1:5000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-805 detects FoxO3.
PA1-805 has been successfully used in Western blot procedures. In Western blot analysis of rat heart and lung, this antibody detects a ~ 71 kDa protein representing FoxO3.
The PA1-805 immunogen is a synthetic peptide corresponding to residues L N(654) V G N F T G A K Q A S S Q S W V(670) of human FOXO3. This sequence is conserved in human, mouse, and rat.
FoxO (forkhead box O) proteins are a family of transcription factors characterized by the distinct forkhead domain. The FoxO proteins act in the insulin signaling pathway, but FoxO3 has also been shown to play a role in the regulation of apoptosis, cell cycle progression and cancer. Several splice variants of FoxO3 have been reported.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.