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Immunofluorescence analysis of cJUN was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with cJUN Mouse Monoclonal Antibody (397500) at 1 µg/mL and incubated for 3 hours at room temperature and then labeled with Alexa Flour 488 Rabbit Anti-Mouse IgG Secondary Antibody (A11059) at a dilution of 1:400 for 30 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 594 Phalloidin (A12381). Panel d is a merged image showing nuclear localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Rabbit , Human , Mouse|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Recombinant protein encompassing an internal region of the human c-Jun protein|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||3-5 ug x 10^6 cells|
|Immunocytochemistry (ICC)||1-2 ug/ml|
|Immunofluorescence (IF)||1-2 ug/ml|
|Western Blot (WB)||1-3 ug/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.
Fos and Jun dimerize to form Activator Protein-1 (AP-1), a transcriptional factor that binds to the 12-O-tetradecanoylphorbol 13-acetate (TPA) response element (TRE) of several cellular and viral genes including human collagenase, metallothionein IIa, stromelysin, interleukin 2, SV40 and polyoma. Fos and Jun contain the ‘leucine-zipper"e; motif that allows for dimerization and an adjacent basic domain required for biological activity. The functionally active form of Fos is in a heterodimer with a member of the Jun family. While Jun family members can form functional homodimers, studies indicate that Fos family members do not self-associate and therefore do not bind DNA on their own. The various dimers differ in their ability to transactivate AP-1 dependent genes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Junc, c-Jun, AP1, AP-1, c-jun
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