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Immunofluorescence analysis of FOXL2 was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with FOXL2 Rabbit Polyclonal Antibody (PA1802) 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||Hamster, Human, Mouse|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide corresponding to residues M(1) M A S Y P E P E D T A G T(14) of mouse FoxL2.|
|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 *|
|ChIP assay (ChIP)||1-3 µg x 10^6 cells|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||0.2 µg/ml|
|Western Blot (WB)||0.2 µg/ml|
* 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.
|Western Blot (WB)||See 1 publications below|
PA1-802 detects FoxL2 protein in mouse and hamster samples.
PA1-802 has successfully been used in Western blot , immunofluorescence and immunohistochemistry procedures. By Western blot, this antibody detects an ~39 kDa protein representing FozL2 in CHO whole cell lysate.
The PA1-802 immunogen is a synthetic Peptide corresponding to residues M(1) M A S Y P E P E D T A G T(14) of mouse FoxL2. This sequence is completely conserved in mouse and rat and 92% conserved in human. This peptide (Cat. # PEP-238) is available for use in neutralization and control experiments.
FOXL2 is a winged helix/forkhead transcription factor that is highly conserved in human, goat, mouse, and certain aquatic species. Given its cross-species conservation, it appears to play a critical role evolutionarily, likely in ovarian differentiation in mammals and in ovarian somatic cell differentiation and in further follicle development and/or maintenance. Immunohistochemical data shows that FOXL2 is a nuclear protein specifically expressed in eyelids and in fetal and adult ovarian follicular cells. It does not undergo any major posttranslational modifications.
FOXL2 is also found to be associated with blepharo-phimosis/ptosis/epicanthus inverses syndrome (BPES). There are two forms of BPES. Type I BPES shows eyelid abnormal-ities that are associated with ovarian failure. In type II BPES, only eyelid defects are found. Studies have shown that FOXL2 is expressed in the mesenchyme of developing mouse eyelids and in adult ovarian follicles
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Foxl2, a forkhead transcription factor, modulates nonclassical activity of the estrogen receptor-alpha.
PA1-802 was used in western blot to study the role of a forkhead transcription factor Foxl2 in the regulation of estrogen receptor alpha activity.
|Kim SY,Weiss J,Tong M,Laronda MM,Lee EJ,Jameson JL||Endocrinology (150:5085)||2009|
forkhead box L2; forkhead box protein L2; forkhead transcription factor FOXL2; P-Frk; pituitary forkhead factor; putative transcription factor foxl2
AU045128; BPES; BPES1; FOXL2; PFRK; PINTO; POF3