Name: Anti-IRF5 antibody [EPR17067]
Brand: RabMAb
SKU: ab181553
Rating: 3 (2 reviews)

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Western blot - Anti-IRF5 antibody [EPR17067] (ab181553)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR17067] to IRF5
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 647 Carrier Free

Product name

Anti-IRF5 antibody [EPR17067]
See all IRF5 primary antibodies
Description

Rabbit monoclonal [EPR17067] to IRF5
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: IRF5 transfected HEK293T whole cell lysate; Ramos, THP-1, A20 and RAW 264.7 whole cell lysates; Mouse thymus, rat thymus, mouse spleen and rat spleen lysates. IHC-P: Human spleen, Human breast cancer, Human lung cancer, Human tonsil, mouse liver and rat spleen tissues. ICC/IF: RAW 264.7 and THP-1 cells. Flow Cyt (intra): THP-1 cells. IP: RAW 264.7 whole cell lysate
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR17067
Isotype

IgG
Research areas
- Neuroscience
- Processes

Alternative Versions
Compatible Secondaries
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human IRF5 protein (ab114295)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab181553 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB		1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
IHC-P	(1)	1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/1000.
IP		1/150.

Notes
Flow Cyt (Intra) 1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB 1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/1000.
IP 1/150.

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.
Target information above from: UniProt accession Q13568 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3663 Human
- Entrez Gene: 27056 Mouse
- Entrez Gene: 296953 Rat
- Omim: 607218 Human
- SwissProt: Q13568 Human
- SwissProt: P56477 Mouse
- Unigene: 521181 Human
- Unigene: 6479 Mouse
Alternative names
- Interferon regulatory factor 5 antibody
- Interferon regulatory factor 5 bone marrow variant antibody
- IRF 5 antibody
- IRF-5 antibody
- Irf5 antibody
- IRF5_HUMAN antibody
- SLEB10 antibody
see all

Western blot - Anti-IRF5 antibody [EPR17067] (ab181553)

All lanes : Anti-IRF5 antibody [EPR17067] (ab181553) at 1/2000 dilution

Lane 1 : Mouse thymus lysate
Lane 2 : Rat thymus lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 55 kDa
Observed band size: 55 kDa

Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-IRF5 antibody [EPR17067] (ab181553)

All lanes : Anti-IRF5 antibody [EPR17067] (ab181553) at 1/1000 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: 55 kDa

Lanes 1 - 2: Merged signal (red and green). Green - ab181553 observed at 56 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab181553 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. Ab181553 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] (ab181553)

Anti-IRF5 antibody [EPR17067] (ab181553)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling IRF5 with ab181553 at a dilution of 1:500. Heat mediated antigen retrieval was performed using AR9 antigen retrieval solution, and microwave treatment for 15 min at 20% power. Anti-Rabbit/Mouse HRP polymer (PerkinElmer Opal Polymer HRP Ms Plus Rb) was used as secondary antibody. Opal tyramide amplification was performed using Opal 520 fluorophore. Counterstained with DAPI stain. Image scanned with Vectra 3.0 and analyzed via Phenochart software.
This image was courteously provided by Dr. Houssein Abdul Sater, Georgia Cancer Center.
Immunocytochemistry/ Immunofluorescence - Anti-IRF5 antibody [EPR17067] (ab181553)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized THP-1 (Human monocytic leukemia cells) cells labeling IRF5 with ab181553 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining on THP-1 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab181553 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-IRF5 antibody [EPR17067] (ab181553)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) cells labeling IRF5 with ab181553 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining on RAW 264.7 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab181553 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Western blot - Anti-IRF5 antibody [EPR17067] (ab181553)

All lanes : Anti-IRF5 antibody [EPR17067] (ab181553) at 1/5000 dilution

Lane 1 : Non-transfected HEK293T (Human epithelial cells from embryonic kidney) whole cell lysate
Lane 2 : IRF5 transfected HEK293T (Human epithelial cells from embryonic kidney) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 55 kDa
Observed band size: 55 kDa

Exposure time: 1 minute

Transfected lysate from abcam (Cat#:ab94265).
Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-IRF5 antibody [EPR17067] (ab181553)

Anti-IRF5 antibody [EPR17067] (ab181553) at 1/2000 dilution + Ramos (Human Burkitt's lymphoma cell line) whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 55 kDa
Observed band size: 55 kDa

Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-IRF5 antibody [EPR17067] (ab181553)

All lanes : Anti-IRF5 antibody [EPR17067] (ab181553) at 1/50000 dilution

Lane 1 : THP-1 (Human monocytic leukemia cells) whole cell lysate
Lane 2 : A20 (Mouse reticulum cell sarcoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 55 kDa
Observed band size: 55 kDa

Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-IRF5 antibody [EPR17067] (ab181553)

All lanes : Anti-IRF5 antibody [EPR17067] (ab181553) at 1/2000 dilution

Lane 1 : Mouse spleen lysate
Lane 2 : Rat spleen lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 55 kDa
Observed band size: 55 kDa

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-IRF5 antibody [EPR17067] (ab181553)

Anti-IRF5 antibody [EPR17067] (ab181553) at 1/5000 dilution + RAW 264.7(Mouse macrophage cells transformed with Abelson murine leukemia virus ) whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 55 kDa
Observed band size: 55 kDa

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Flow Cytometry (Intracellular) - Anti-IRF5 antibody [EPR17067] (ab181553)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed THP-1 (Human monocytic leukemia cells) cells labeling IRF5with ab181553 at 1/50 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730;black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] (ab181553)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling IRF5 with ab181553 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and cytoplasmic staining on leukocytes of Human spleen is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] (ab181553)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling IRF5 with ab181553 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and cytoplasmic staining on leukocyte of Human breast cancer stroma is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] (ab181553)

Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling IRF5 with ab181553 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on cancer cells of Human lung cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] (ab181553)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling IRF5 with ab181553 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on kupffer cells of mouse liver is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] (ab181553)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling IRF5 with ab181553 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and cytoplasmic staining on lymphocytes of rat spleen is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-IRF5 antibody [EPR17067] (ab181553)

IRF5 was immunoprecipitated from 1mg of RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate with ab181553 at 1/150 dilution. Western blot was performed from the immunoprecipitate using ab181553 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: RAW 264.7 whole cell lysate 10 µg (Input).
Lane 2: ab181553 IP in RAW 264.7 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab181553 in RAW 264.7 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Anti-IRF5 antibody [EPR17067] (ab181553)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

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Publishing research using ab181553? Please let us know so that we can cite the reference in this datasheet.

ab181553 has been referenced in 12 publications.

Yin X et al. MDA5 Governs the Innate Immune Response to SARS-CoV-2 in Lung Epithelial Cells. Cell Rep 34:108628 (2021). PubMed: 33440148
Motani K & Kosako H BioID screening of biotinylation sites using the avidin-like protein Tamavidin 2-REV identifies global interactors of stimulator of interferon genes (STING). J Biol Chem 295:11174-11183 (2020). PubMed: 32554809
Konishi T et al. Fibrotic liver has prompt recovery after ischemia-reperfusion injury. Am J Physiol Gastrointest Liver Physiol 318:G390-G400 (2020). PubMed: 31961717
Heinz LX et al. TASL is the SLC15A4-associated adaptor for IRF5 activation by TLR7-9. Nature 581:316-322 (2020). PubMed: 32433612
Bian X et al. The absence of oestrogen receptor beta disturbs collagen I type deposition during Achilles tendon healing by regulating the IRF5-CCL3 axis. J Cell Mol Med 24:9925-9935 (2020). PubMed: 32776630

View all Publications for this product

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1-2 of 2 Abreviews or Q&A

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (lung)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: 10 mM sodium citrate

Permeabilization

Specification

lung

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 07 2020

Application

ChIP

Sample

Mouse Cell lysate - nuclear (LPS-stimulated bone marrow-derived macrophages)

Negative control

non-binding regions, e.g. Arg1 gene body

Specification

LPS-stimulated bone marrow-derived macrophages

Detection step

Real-time PCR

Type

Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde

Positive control

same sample, using different Ab, selected based on previous ChIP-seq data sets

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 04 2017

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (12)

Customer reviews and Q&As

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-IRF5 antibody [EPR17067]

ChIP abreview for Anti-IRF5 antibody [EPR17067]