The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 39 kDa.
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use a concentration of 10 µg/ml.
FunctionSpecifically binds to the upstream regulatory region of type I IFN and IFN-inducible MHC class I genes (the interferon consensus sequence (ICS)) and represses those genes. Also acts as an activator for several genes including H4 and IL7. Constitutively binds to the ISRE promoter to activate IL7. Involved in cell cycle regulation through binding the site II (HiNF-M) promoter region of H4 and activating transcription during cell growth. Antagonizes IRF1 transcriptional activation.
Tissue specificityExpressed throughout the epithelium of the colon. Also expressed in lamina propria.
Sequence similaritiesBelongs to the IRF family. Contains 1 IRF tryptophan pentad repeat DNA-binding domain.
Post-translational modificationsAcetylated by CBP/ p300 during cell-growth. Acetylation on Lys-75 is required for stimulation of H4 promoter activity. The major sites of sumoylation are Lys-137 and Lys-293. Sumoylation by SUMO1 increases its transcriptional repressor activity on IRF1 and diminishes its ability to activate ISRE and H4 promoter.
Lane 1: Wild type HAP1 whole cell lysate (20 µg) Lane 2: IRF2 knockout HAP1 whole cell lysate (20 µg) Lane 3: HeLa whole cell lysate (20 µg) Lane 4: CACO2 whole cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab55331 observed at 48 kDa. Red - loading control, ab176560, observed at 50 kDa.
ab55331 was shown to specifically react with IRF2 when IRF2 knockout samples were used. Wild-type and IRF2 knockout samples were subjected to SDS-PAGE. Ab55331 and ab176560 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - IRF2 antibody (ab55331)
Predicted band size : 39 kDa IRF2 antibody (ab55331) at 1ug/lane + HeLa cell lysate at 25ug/lane.
IHC image of ab55331 staining in human colonic carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab55331, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
References for Anti-IRF2 antibody (ab55331)
This product has been referenced in:
Li Y et al. Inflammatory signaling regulates embryonic hematopoietic stem and progenitor cell production. Genes Dev28:2597-612 (2014).
Read more (PubMed: 25395663) »
Larrea E et al. Dysregulation of interferon regulatory factors impairs the expression of immunostimulatory molecules in hepatitis C virus genotype 1-infected hepatocytes. GutN/A:N/A (2013).
Read more (PubMed: 23787026) »