Immunofluorescence analysis of JunB was done on 70% confluent log phase PC-3 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 JunB Rabbit Polyclonal Antibody (PA1835) 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|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues D(45) P Y R G L K G P G A R G P G P E(61) of mouse JunB.|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1-3 ul|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||10 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.
PA1-835 detects JunB from mouse cells.
PA1-835 has been successfully used in ChiP, Western blot and immunoprecipitation procedures. By Western blot, this antibody detects an ~38 kDa protein representing JunB from both uninduced and induced L929 cells.
The PA1-835 immunogen is a synthetic peptide corresponding to residues D(45) P Y R G L K G P G A R G P G P E(61) of mouse JunB.
Reconstitute with PBS.
Cellular oncogenes, or proto-oncogenes, play pivotal roles in cellular communication pathways that regulate normal growth, development and differentiation. The cellular oncogene families fos and jun encode nuclear proteins that can function as transcription factors. The fos family of nuclear oncogenes encode cFos, FosB, (fos-related antigen) Fra1, and Fra2.
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 and quote;leucine-zipper and quote; 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.
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