The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC - Wholemount
1/100 - 1/500. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
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.
Anti-c-Jun antibody (ab86740) at 1/500 dilution + 35µg of HEK293 whole cell lysate
Predicted band size: 36 kDa Observed band size: 36 kDa Additional bands at: 30 kDa. We are unsure as to the identity of these extra bands.
IHC - Wholemount - Anti-c-Jun antibody (ab86740)This image is courtesy of an anonymous Abreview
ab86740 staining 24hpf WT zebrafish embryos by IHC-wholemount. The embryos were dechorionated and fixed with 4% paraformaldehyde (or Dent's fixative) over 48h at 4 °C. Embryos were then washed two times with PBS and two times with MeOH, permeabilized with methanol for 5 minutes and blocked with 10% goat serum in PBS - Triton for 1 hour. Staining with ab86740 at a 1/100 dilution in PBS-10% goat serum- Triton was performed for 22h at 4°C. A goat anti-rabbit Alexa 568 polyclonal antibody at 1/1000 was used as the secondary antibody.