Anti-IRF5 antibody [10T1] (ab33478)
Key features and details
- Mouse monoclonal [10T1] to IRF5
- Suitable for: ICC/IF, WB, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG1
Overview
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Product name
Anti-IRF5 antibody [10T1]
See all IRF5 primary antibodies -
Description
Mouse monoclonal [10T1] to IRF5 -
Host species
Mouse -
Specificity
ab33478 recognises IRF-5. -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF MouseHumanWB MouseHuman -
Immunogen
Recombinant human IRF-5 (176-240 aa, isoform D) purified from E. coli. NCBI Accession No.: NP_001092099 (formally NP_002191).
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Positive control
- WB: Extracts of Ramos, THP-1 and A20 cells. ICC/IF: THP-1, Raw264.7 and HEK293. Flow Cyt: THP-1 cells.
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General notes
This product was changed from ascites to tissue culture supernatant on 24/01/2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.40
Preservative: 0.1% Sodium azide
Constituent: 99% PBS -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
10T1 -
Myeloma
Sp2/0 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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Flow Cyt |
Human
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ICC/IF |
Mouse
Human
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WB |
Mouse
Human
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Application | Abreviews | Notes |
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ICC/IF |
Use at an assay dependent concentration.
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WB | (1) |
Use at an assay dependent concentration. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
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Flow Cyt |
Use at an assay dependent concentration.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Notes |
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ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa). |
Flow Cyt
Use at an assay dependent concentration. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Target
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Involvement in disease
Genetic 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 similarities
Belongs to the IRF family.
Contains 1 IRF tryptophan pentad repeat DNA-binding domain. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 3663 Human
- Entrez Gene: 27056 Mouse
- Omim: 607218 Human
- SwissProt: Q13568 Human
- SwissProt: P56477 Mouse
- Unigene: 521181 Human
- Unigene: 6479 Mouse
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Alternative names
- Interferon regulatory factor 5 antibody
- Interferon regulatory factor 5 bone marrow variant antibody
- IRF 5 antibody
see all
Images
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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 kDaLanes 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.
This image was produced using the ascites version of this antibody.
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Flow cytometry analysis of IRF5 in THP-1 cell line, staining at 2-5µg for 1x106 cells (red line). The secondary antibody used goat anti-mouse IgG Alexa fluor 488 conjugate. Isotype control antibody was mouse IgG (black line).
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Immunocytochemistry/ Immunofluorescence analysis of IRF5 in Raw264.7 cells. The cell was stained with ab33478 (1:100). The secondary antibody (green) was used Alexa Fluor 488. DAPI was stained the cell nucleus (blue).
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Immunocytochemistry/ Immunofluorescence analysis of IRF5 in THP-1 cells. The cell was stained with ab33478 (1:100). The secondary antibody (green) was used Alexa Fluor 488. DAPI was stained the cell nucleus (blue).
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Anti-IRF5 antibody [10T1] (ab33478) at 1/1000 dilution + THP-1 cell lysate
Secondary
Goat anti-mouse secondary antibody conjugated to HRP
Predicted band size: 58 kDa -
Immunofluorescence/Immunocytochemistry analysis of HEK293 cells (105-106) stained with Mouse monoclonal [10T1] to IRF5 (ab33478 1/100-1/200).
This image was produced using the ascites version of this antibody.
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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
Secondary
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.
This image was produced using the ascites version of this antibody.
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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.
This image was produced using the ascites version of this antibody.
Protocols
References (14)
ab33478 has been referenced in 14 publications.
- Moen SH et al. Human Toll-like Receptor 8 (TLR8) Is an Important Sensor of Pyogenic Bacteria, and Is Attenuated by Cell Surface TLR Signaling. Front Immunol 10:1209 (2019). PubMed: 31214180
- 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). PubMed: 28620671
- Koelzer VH et al. Digital analysis and epigenetic regulation of the signature of rejection in colorectal cancer. Oncoimmunology 6:e1288330 (2017). PubMed: 28507795
- Chen X et al. MicroRNA-302a suppresses influenza A virus-stimulated interferon regulatory factor-5 expression and cytokine storm induction. J Biol Chem 292:21291-21303 (2017). PubMed: 29046356
- Li D et al. Specific detection of interferon regulatory factor 5 (IRF5): A case of antibody inequality. Sci Rep 6:31002 (2016). PubMed: 27481535