Western blot analysis of FOXO1 was performed by loading 25 ug of Hela (lane 1), 293T (lane 2) and rat heart (lane 3) onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a FOXO1 polyclonal antibody (Product # PA1-804) at a dilution of 1:500 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Images were taken at an exposure time of 2 min (lanes 1 and 2) and 1 min (lane 3). Results show a band at ~66kDa.
|Tested species reactivity||Human, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues D(633) N V L P N Q S F P H S V K T(647) of human FOXO1a.|
|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 *|
|Western Blot (WB)||1:100-1:1000|
* 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.
PA1-804 detects FoxO1a in human and rat samples.
PA1- 804 has been successfully used in Western blot procedures. By Western blot this antibody detects a ~66 kDa protein representing FoxO1a in rat heart and lung protein extracts.
The PA1- 804 immunogen is a synthetic peptide corresponding to residues D(633) N V L P N Q S F P H S V K T(647) of human FOXO1a. This sequence is completely conserved in mouse and rat.
Fig. 1 shows a Western blot of FoxO1a in rat lung protein extract using PA1-804.
FoxO1A belongs to the forkhead family of transcription factors, which are involved in many biological processes and function under the control of the insulin signaling pathway. The mammalian DAF-16-like transcription factors, FoxO1A, FkhRL1, and AFX, function as key regulators of insulin signaling, cell cycle progression, and apoptosis downstream of phosphoinositide 3-kinase. Dysfunction in FoxO signaling has been linked to insulin resistance and type II diabetes. The FoxO1A gene is mapped to chromosome 13q14.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.