Product nameAnti-IRF5 antibody [10T1]
See all IRF5 primary antibodies
DescriptionMouse monoclonal [10T1] to IRF5
Specificityab33478 recognises IRF-5.
Tested applicationsSuitable for: ICC/IF, ELISA, IP, WB, Flow Cyt, IHC-Pmore details
Species reactivityReacts with: Mouse, Human
Recombinant human IRF-5 (176-240 aa, isoform D) purified from E. coli. NCBI Accession No.: NP_001092099 (formally NP_002191).
- Extracts of Ramos, THP-1 and A20 cells. HEK293
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.1% Sodium azide
Constituent: 99% PBS
Concentration information loading...
PurityProtein G purified
Light chain typekappa
- Epigenetics and Nuclear Signaling
- Domain Families
- HLH / Leucine Zipper
Our Abpromise guarantee covers the use of ab33478 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||1/100 - 1/200.|
|ELISA||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|WB||1/500 - 1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).|
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-P||1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
Involvement in diseaseGenetic variations in IRF5 are associated with susceptibility to inflammatory bowel disease type 14 (IBD14) [MIM:612245]. IBD14 is a chronic, relapsing inflammation of the gastrointestinal tract with a complex etiology. It is subdivided into Crohn disease and ulcerative colitis phenotypes. Crohn disease may affect any part of the gastrointestinal tract from the mouth to the anus, but most frequently it involves the terminal ileum and colon. Bowel inflammation is transmural and discontinuous; it may contain granulomas or be associated with intestinal or perianal fistulas. In contrast, in ulcerative colitis, the inflammation is continuous and limited to rectal and colonic mucosal layers; fistulas and granulomas are not observed. Both diseases include extraintestinal inflammation of the skin, eyes, or joints.
Genetic variations in IRF5 are associated with susceptibility to systemic lupus erythematosus type 10 (SLEB10) [MIM:612251]. Systemic lupus erythematosus (SLE) is a chronic, inflammatory and often febrile multisystemic disorder of connective tissue. It affects principally the skin, joints, kidneys and serosal membranes. It is thought to represent a failure of the regulatory mechanisms of the autoimmune system.
Genetic variations in IRF5 are a cause of susceptibility to rheumatoid arthritis (RA) [MIM:180300]. It is a systemic inflammatory disease with autoimmune features and a complex genetic component. It primarily affects the joints and is characterized by inflammatory changes in the synovial membranes and articular structures, widespread fibrinoid degeneration of the collagen fibers in mesenchymal tissues, and by atrophy and rarefaction of bony structures.
Sequence similaritiesBelongs to the IRF family.
Contains 1 IRF tryptophan pentad repeat DNA-binding domain.
- Information by UniProt
- Interferon regulatory factor 5 antibody
- Interferon regulatory factor 5 bone marrow variant antibody
- IRF 5 antibody
All lanes : Anti-IRF5 antibody [10T1] (ab33478) at 1/500 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : IRF5 knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 58 kDa
Lanes 1 - 2: Merged signal (red and green). Green - ab33478 observed at 56 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab33478 was shown to specifically react with IRF5 in wild-type HAP1 cells as signal was lost in IRF5 knockout cells. Wild-type and IRF5 knockout samples were subjected to SDS-PAGE. Ab33478 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Immunofluorescence/Immunocytochemistry analysis of HEK293 cells (105-106) stained with Mouse monoclonal [10T1] to IRF5 (ab33478 1/100-1/200).
All lanes : Anti-IRF5 antibody [10T1] (ab33478) at 1/1000 dilution
Lane 1 : Ramos cell lysate
Lane 2 : THP-1 cell lysate
Lane 3 : A20 cell lysate
Lane 4 : NIH 3T3 cell lysate
All lanes : goat anti-mouse secondary antibody conjugated to HRP
Developed using the ECL technique.
Predicted band size: 58 kDa
Observed band size: 58 kDa
Additional bands at: 26 kDa, 80 kDa. We are unsure as to the identity of these extra bands.
100-200µg of whole cell lysates were used for western blotting.
Overlay histogram showing THP1 cells stained with ab33478 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab33478, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in THP1 cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
ab33478 at 1/50 dilution staining IRF5 in Human normal colon mucosa tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). The section was incubated with the primary antibody for 2 hours at room temperature. Antigen retrieval was performed in 0.1M sodium citrate buffer and detected using Diaminobenzidine (DAB).
ab33478 at 1/50 dilution staining IRF5 in Human colon adenocarcinoma tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). The section was incubated with the primary antibody for 2 hours at room temperature. Antigen retrieval was performed in 0.1M sodium citrate buffer and detected using Diaminobenzidine (DAB).
This product has been referenced in:
- Xiao Y et al. An oligodeoxynucleotide with AAAG repeats significantly attenuates burn-induced systemic inflammatory responses via inhibiting interferon regulatory factor 5 pathway. Mol Med 23:N/A (2017). Read more (PubMed: 28620671) »
- Koelzer VH et al. Digital analysis and epigenetic regulation of the signature of rejection in colorectal cancer. Oncoimmunology 6:e1288330 (2017). Read more (PubMed: 28507795) »