ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
Transcription factor that recognizes and binds to the enhancer heptamer motif 5'-TGA[CG]TCA-3'. Promotes activity of NR5A1 when phosphorylated by HIPK3 leading to increased steroidogenic gene expression upon cAMP signaling pathway stimulation. Involved in activated KRAS-mediated transcriptional activation of USP28 in colorectal cancer (CRC) cells (PubMed:24623306). Binds to the USP28 promoter in colorectal cancer (CRC) cells (PubMed:24623306).
Belongs to the bZIP family. Jun subfamily. Contains 1 bZIP (basic-leucine zipper) domain.
Ubiquitinated by the SCF(FBXW7), leading to its degradation. Ubiquitination takes place following phosphorylation, that promotes interaction with FBXW7. Phosphorylated by CaMK4 and PRKDC; phosphorylation enhances the transcriptional activity. Phosphorylated by HIPK3. Phosphorylated by DYRK2 at Ser-243; this primes the protein for subsequent phosphorylation by GSK3B at Thr-239. Phosphorylated at Thr-239, Ser-243 and Ser-249 by GSK3B; phosphorylation reduces its ability to bind DNA. Phosphorylated by PAK2 at Thr-2, Thr-8, Thr-89, Thr-93 and Thr-286 thereby promoting JUN-mediated cell proliferation and transformation. Phosphorylated by PLK3 following hypoxia or UV irradiation, leading to increase DNA-binding activity. Acetylated at Lys-271 by EP300.
ChIP - Anti-c-Jun antibody - ChIP Grade (ab31419)This image is courtesy of an anonymous abreview.
ChIP analysis using ab31419 binding c-Jun in human endothelial cells (EA.hy926). Cells were cross-linked for 10 minutes with formaldehyde then incubated with undiluted primary antibody for 10 hours at 4°C in 1x ChIP buffer. Protein binding was detected using real-time PCR. Positive control: Position 89340150-89340297 in chromosome 11 (has a validated c-Jun site). Negative Control: Igr5 intron 3 (contains no c-Jun binding site).
ICC/IF image of HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling c-Jun (green) with ab31419 at 1 µg/ml. The cells were fixed in 4% PFA (10 minutes) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with ab31419 at 1 µg/ml overnight at +4 °C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) ab150077 used at a 1/1000 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.
Western blot - Anti-c-Jun antibody - ChIP Grade (ab31419)
All lanes : Anti-c-Jun antibody - ChIP Grade (ab31419) at 1/500 dilution
Lane 1 : Extracts of Hela (Human epithelial cell line from cervix adenocarcinoma) cells Lane 2 : Extracts of Hela (Human epithelial cell line from cervix adenocarcinoma) cells with immunizing peptide
Flow Cytometry - Anti-c-Jun antibody - ChIP Grade (ab31419)This image is courtesy of an anonymous abreview.
Overlay histogram showing Jurkat (Human T cell leukemia cell line from peripheral blood) cells stained with ab31419 (blue line) at a 1/300 dilution. The cells were prepared as follows: Spin down, wash in FACS buffer 1x, fix, wash 2x, and stain with primary antibody overnight. Buffer was 0.1% sodium azide with FBS in phosphate buffered solution. Cells were fixed using paraformaldehyde and permeabilized using Triton X-100 and NP40. Cells were gated by isolating cell population from plot of SSC-A / FSA-A. The secondary antibody used was a FITC-conjugated Goat anti-Rabbit polyclonal, diluted 1/100.