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Immunofluorescence analysis of Phospho-c-Fos pThr325 was done on 70% confluent log phase HeLa cell treated with 200 nM of PMA for 20 minutes. The cell 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 Phospho-c-Fos pThr325 Rabbit Polyclonal Antibody (44281G) at 2 ug/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 and cytoplasmic localization. Panel e is untreated cell with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human c-Fos that contains threonine 325. The sequence is conserved in human, mouse and rat.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1-3 µg x 10^6 cells|
|Immunocytochemistry (ICC)||2-3 µg/ml|
|Immunofluorescence (IF)||2-3 µg/ml|
|Western Blot (WB)||Assay Dependent|
* 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.
c-FOS is a nuclear phosphoprotein which forms a tight but non-covalently linked complex with the JUN/AP-1 transcription factor. It has a critical function in regulating the development of cells destined to form and maintain the skeleton, and is thought to have an important role in signal transduction, cell proliferation and differentiation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
activator protein 1; cellular oncogene c-fos; FBJ murine osteosarcoma viral (v-fos) oncogene homolog (oncogene FOS); FBJ murine osteosarcoma viral oncogene homolog; FOS; Fos proto-oncogene, AP-1 trancription factor subunit; G0/G1 switch regulatory protein 7; G0S7
AP-1; C-FOS; FOS; G0S7; p55