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 Carrier Free

产品名称

Anti-IRF5抗体[EPR17067]
参阅全部 IRF5 一抗
描述

兔单克隆抗体[EPR17067] to IRF5
宿主

Rabbit
经测试应用

适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details
种属反应性

与反应: Mouse, Rat, Human
免疫原

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
阳性对照
- 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
常规说明
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.

形式

Liquid
存放说明

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.
存储溶液

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

Protein A purified
克隆

单克隆
克隆编号

EPR17067
同种型

IgG
研究领域
- Neuroscience
- Processes

Alternative Versions
- Alexa Fluor® 488 Anti-IRF5 antibody [EPR17067] (ab208271)
- Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
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

Abpromise™承诺保证使用ab181553于以下的经测试应用

“应用说明”部分下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用	Ab评论	说明
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.

说明
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.

疾病相关

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.
序列相似性

Belongs to the IRF family.
Contains 1 IRF tryptophan pentad repeat DNA-binding domain.
细胞定位

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
数据库链接
- 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
别名
- 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 1/1000 dilution

Lane 1 : Wild-type A549 cell lysate
Lane 2 : IRF5 knockout A549 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 55 kDa
Observed band size: 50-70 kDa why is the actual band size different from the predicted?

Western blot: Anti-IRF5 antibody [EPR17067] (ab181553) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab181553 was shown to bind specifically to IRF5. A band was observed at 50-70 kDa in wild-type A549 cell lysates with no signal observed at this size in IRF5 knockout cell line. To generate this image, wild-type and IRF5 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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

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发表研究结果有使用 ab181553？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab181553 被引用在 24 文献中.

Boeszoermenyi A et al. A conformation-locking inhibitor of SLC15A4 with TASL proteostatic anti-inflammatory activity. Nat Commun 14:6626 (2023). PubMed: 37863876
Chen X et al. Structural basis for recruitment of TASL by SLC15A4 in human endolysosomal TLR signaling. Nat Commun 14:6627 (2023). PubMed: 37863913
Li D et al. Pathogenic Tconvs promote inflammatory macrophage polarization through GM-CSF and exacerbate abdominal aortic aneurysm formation. FASEB J 36:e22172 (2022). PubMed: 35133017
Xia W et al. Excess fatty acids induce pancreatic acinar cell pyroptosis through macrophage M1 polarization. BMC Gastroenterol 22:72 (2022). PubMed: 35183119
Qiao X et al. Interferon regulatory factor 5‑induced upregulation of zinc‑finger protein 217 promotes pancreatic carcinoma progression. Mol Med Rep 25:N/A (2022). PubMed: 35362545

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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

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提交于 Jan 07 2020

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Key features and details

Related conjugates and formulations

概述

产品名称

描述

宿主

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

应用

The Abpromise guarantee

靶标

疾病相关

序列相似性

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

SDS download

Datasheet download

Certificate of Compliance

文献 (24)

客户评价及客户问答

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