Invitrogen

IRF4 Monoclonal Antibody (3E4), eBioscience™

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

22 Published Figures

21 References

View all (33) IRF4 antibodies

Datasheet

Protocols

Questions & Answers

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

IRF4 Antibody in Western Blot (WB) — Western blot was performed using Anti-IRF4 Monoclonal Antibody (3E4) (Product # 14-9858-82) and a 51kDa band corresponding to IRF4 was observed in IM9 and was absent in MOLT-4, Jurkat and U-937 as reported. Whole cell extracts (30 µg lysate) of IM9 (Lane 1), MOLT-4 (Lane 2), Jurkat (Lane 3) and U-937 (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (5ug/ml) and detected by chemiluminescence F(ab)2-Rabbit anti-Rat IgG (H+L) Secondary Antibody, HRP (Product # PA1-29927; 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

IRF4 Antibody in Immunohistochemistry (IHC) — Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Immunohistochemistry

IRF4 Antibody in Flow Cytometry (Flow) — Figure 4. B cells promote CD8 T cell memory programming following subunit vaccination (A) Representative histograms for Eomes, CD122, IRF4, and TCF1 staining of tetramer+ cells from WT (black) or MD4 (maroon) mice 7 days after combined-adjuvant vaccination or for CD44- naive control CD8 T cells (gray). (B) The gMFI of tetramer+ cells for Eomes, CD122, and IRF4, where the x axis intersects the y axis at the average gMFI value for CD44- naive controls, and the percentage of tetramer+ cells that stained positive for TCF1. (C and D) Representative KLRG1 versus CD127 staining (C) and numbers of tetramer+ splenocytes in the respective quadrants (D), 7 days after immunization. Data shown are means +- SEM; n : mice per group, representative of 3 experiments. Significance was defined by two-tailed, unpaired Student's t tests, where *p < 0.05, **p < 0.01, and ***p < 0.001.

IRF4 Antibody in ChIP assay (ChIP) — Figure 3. Increased Binding of IRF4 at the Il17 Locus and IRF4 and RORgammat Requirement for IL-17A Production in Blimp-1 -/- Foxp3 + Treg (A) Representation of the Il17 locus showing RORgammat and IRF4 consensus binding sites (top) and RORgt ChIP (bottom) in Blimp-1 +/+ and Blimp-1 -/- CD4 + Foxp3 GFP+ Treg and wild-type pTh17 cells. (B) IL-17A expression in TCRbeta + CD4 + Foxp3 + Treg (MLNs) from Prdm1 +/+ Rorc +/+ Foxp3 CREYFP+ (Ctrl), Prdm1 F/F Rorc +/+ Foxp 3CREYFP+ (knockout [KO]), and Prdm1 F/F Rorc F/F Foxp3 CREYFP+ (double knockout [DKO]) mice and expression of Foxp3 , Prdm1 , Rorc , and Il17a mRNA (bottom graphs) in sorted and in vitro -stimulated (phorbol myristate acetate [PMA] and ionomycin for 4 hr) CD4 + Foxp3 CREYFP+ cells. (C) IRF4 ChIP at the Il17 CNS7 and Il10 Intron I regions in Blimp1 +/+ and Blimp1 -/- CD4 + Foxp3 GFP+ Treg and wild-type pTh17 cells. (D) Irf4 , Il17a , and Il10 mRNA expression following Irf4 siRNA knockdown (SC, scrambled control). Data shown are representative of at least two independent experiments. Error bars show SEM (n >= 3 per group). *p < 0.05 and **p < 0.01, one-way ANOVA.

IRF4 Antibody in ELISA (ELISA) — Figure 2. SphK1 -/- T Cells Secrete Both IFN-gamma and IL-17 upon TCR Stimulation (A-C) pMel and pMel-SphK -/- T cells activated for 3 days with cognate antigen gp100 were antigen-restimulated overnight, and the supernatant was used for ELISA-based determination of secreted cytokine: (A) IFN-gamma, (B) IL-17, (C) RORgammat and IRF4 levels. (D) Splenic T cells from IL-17-GFP reporter or SphK1 -/- -IL-17-GFP mice were cultured in Th17 polarizing condition for 3 days, restimulated with anti-CD3 (2 ug/mL) and anti-CD28 antibody (5 ug/mL) for 4 h, and the percentage of IL-17-GFP-positive cells were determined by flow cytometry. (E) Three-day TCR-activated WT and SphK1 -/- T cells were analyzed for expression of stemness-associated genes using mRNA. Data are representative of three independent experiments in (A)-(E). (F) C57BL/6 mice (n = 10 mice/group) were inoculated subcutaneously (s.c.) with 0.3 x 10 6 B16-F10 melanoma cells for 9 days, after which mice were either kept untreated or adoptively transferred, either with 1 x 10 6 pMel or pMel-SphK1 -/- T cells. Tumor growth was measured using digital calipers twice weekly. Left panel: estimated growth curves for each experimental condition. Right panel: survival curve (p value pMel versus pMel SphK1 -/- is 1e-05). (G) Upper panel: C57BL/6 mice (n = 5 mice/group) were inoculated (s.c.) with 0.25 x 10 6 B16-F10 melanoma cells for 14 days, then 1 x 10 6 3 days gp100-activated pMel T cells, pMel SphK1 -/- T cells, or pMel T cells trea

IRF4 Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell lines tested owing to their inherent genetic constitution. Relative expression of IRF4 was observed in IM9 in comparison to MOLT-4, Jurkat and U-937 using Anti-IRF4 Monoclonal antibody (3E4) (Product # 14-9858-82) in Western Blot. {RE}

Product Details

14-9858-82

Applications	Tested Dilution	Publications
Western Blot (WB)	5 µg/mL	View 1 publication 1 publication
Immunohistochemistry (IHC)	-	View 1 publication 1 publication
Flow Cytometry (Flow)	-	View 17 publications 17 publications
ELISA (ELISA)	-	View 1 publication 1 publication
ChIP assay (ChIP)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Mouse
Published species	Human, Mouse
Host/Isotype	Rat / IgG1, kappa
Class	Monoclonal
Type	Antibody
Clone	3E4
Conjugate	Unconjugated Alexa Fluor 700 eFluor 450 eFluor 660 FITC PE PE-Cyanine7 PE-eFluor 610 PerCP-eFluor 710
Form	Liquid
Concentration	0.5 mg/mL
Purification	Affinity chromatography
Storage buffer	PBS, pH 7.2
Contains	0.09% sodium azide
Storage conditions	4° C
RRID	AB_10804654

Product Specific Information

Description: The monoclonal antibody 3E4 reacts with human and mouse interferon regulatory factor 4 (IRF4). IRF4 is a 52 kDa transcription factor with roles in B cell, T cell and macrophage function. In B cells, IRF4 is highly expressed in mature plasma cells and plays a crucial role in their differentiation. IRF4 has been shown to interact with PU.1 and control the transcription of many B cell-specific genes including Prdm1, which encodes Blimp1. In T cells, IRF4 has been implicated in regulatory T (Treg), T helper (Th)2, Th9 and Th17 cell development and function. This transcription factor is upregulated upon T cell activation and is expressed in mature T cells. Studies have shown that IRF4 directly induces Blimp1 expression in Tregs, leading to IL-10 expression. IRF4 has also been demonstrated to be involved in macrophage polarization and regulation. Lastly, in addition to its roles in normal immune function, IRF4 expression has been reported to be upregulated in many blood-related cancers.

Applications Reported: This 3E4 antibody has been reported for use in western blotting.

Applications Tested: This 3E4 antibody has been tested by western blot of reduced cell lysate prepared from splenocytes cultured for 3 days in the presence of plate-bound Anti-Mouse CD3 and soluble Anti-Mouse CD28. This antibody can be used at less than or equal to 5 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Purity: Greater than 90%, as determined by SDS-PAGE.

Aggregation: Less than 10%, as determined by HPLC.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

The IRF (interferon regulatory factor) family of transcription factors are characterized by an unique tryptophan pentad repeat DNA-binding domain. The IRFs are important in the regulation of interferons in response to infection by virus, and in the regulation of interferon-inducible genes. IRF4 is lymphocyte specific and negatively regulates toll-like-receptor (TLR) signaling that is central to the activation of innate and adaptive immune systems. A chromosomal translocation involving this gene and the IgH locus, t(6;14)(p25;q32), may be a cause of multiple myeloma. Alternatively spliced transcript variants have been found.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

CD4⁺ T Cell Fate Decisions Are Stochastic, Precede Cell Division, Depend on GITR Co-Stimulation, and Are Associated With Uropodium Development.

Frontiers in immunology

Cobbold SP,Adams E,Howie D,Waldmann H

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Mon Jan 31 00:00:00 EST 2022

B cells promote CD8 T cell primary and memory responses to subunit vaccines.

Cell reports

Klarquist J,Cross EW,Thompson SB,Willett B,Aldridge DL,Caffrey-Carr AK,Xu Z,Hunter CA,Getahun A,Kedl RM

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Tue Aug 24 00:00:00 EDT 2021

Mitochondrial Oxidative Phosphorylation Regulates the Fate Decision between Pathogenic Th17 and Regulatory T Cells.

Cell reports

Shin B,Benavides GA,Geng J,Koralov SB,Hu H,Darley-Usmar VM,Harrington LE

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Tue Feb 11 00:00:00 EST 2020

Pro-Survival Lipid Sphingosine-1-Phosphate Metabolically Programs T Cells to Limit Anti-tumor Activity.

Cell reports

Chakraborty P,Vaena SG,Thyagarajan K,Chatterjee S,Al-Khami A,Selvam SP,Nguyen H,Kang I,Wyatt MW,Baliga U,Hedley Z,Ngang RN,Guo B,Beeson GC,Husain S,Paulos CM,Beeson CC,Zilliox MJ,Hill EG,Mehrotra M,Yu XZ,Ogretmen B,Mehrotra S

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in ELISA

Tue Aug 13 00:00:00 EDT 2019

Ufbp1 promotes plasma cell development and ER expansion by modulating distinct branches of UPR.

Nature communications

Zhu H,Bhatt B,Sivaprakasam S,Cai Y,Liu S,Kodeboyina SK,Patel N,Savage NM,Sharma A,Kaufman RJ,Li H,Singh N

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Wed Mar 06 00:00:00 EST 2019

ROCK2, but not ROCK1 interacts with phosphorylated STAT3 and co-occupies TH17/TFH gene promoters in TH17-activated human T cells.

Scientific reports

Chen W,Nyuydzefe MS,Weiss JM,Zhang J,Waksal SD,Zanin-Zhorov A

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Fri Nov 09 00:00:00 EST 2018

Blimp-1 Functions as a Molecular Switch to Prevent Inflammatory Activity in Foxp3⁺RORγt⁺ Regulatory T Cells.

Cell reports

Ogawa C,Bankoti R,Nguyen T,Hassanzadeh-Kiabi N,Nadeau S,Porritt RA,Couse M,Fan X,Dhall D,Eberl G,Ohnmacht C,Martins GA

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in ChIP assay

Tue Oct 02 00:00:00 EDT 2018

RANKL-mediated harmonious dialogue between fetus and mother guarantees smooth gestation by inducing decidual M2 macrophage polarization.

Cell death & disease

Meng YH,Zhou WJ,Jin LP,Liu LB,Chang KK,Mei J,Li H,Wang J,Li DJ,Li MQ

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Thu Oct 12 00:00:00 EDT 2017

Ubiquitin specific peptidase 4 stabilizes interferon regulatory factor protein and promotes its function to facilitate interleukin-4 expression in T helper type 2 cells.

International journal of molecular medicine

Guo Z,Xu P,Ge S,Zhang C,Zheng X,Xu J,Liu Z,Li B,Ge S

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Sun Oct 01 00:00:00 EDT 2017

Ubiquitin specific peptidase 4 stabilizes interferon regulatory factor protein and promotes its function to facilitate interleukin-4 expression in T helper type 2 cells.

International journal of molecular medicine

Guo Z,Xu P,Ge S,Zhang C,Zheng X,Xu J,Liu Z,Li B,Ge S

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Sun Oct 01 00:00:00 EDT 2017

Batf3 selectively determines acquisition of CD8⁺ dendritic cell phenotype and function.

Immunology and cell biology

Chandra J,Kuo PT,Hahn AM,Belz GT,Frazer IH

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Wed Feb 01 00:00:00 EST 2017

Affinity and dose of TCR engagement yield proportional enhancer and gene activity in CD4+ T cells.

eLife

Allison KA,Sajti E,Collier JG,Gosselin D,Troutman TD,Stone EL,Hedrick SM,Glass CK

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Mon Jul 04 00:00:00 EDT 2016

STAT3/5-Dependent IL9 Overexpression Contributes to Neoplastic Cell Survival in Mycosis Fungoides.

Clinical cancer research : an official journal of the American Association for Cancer Research

Vieyra-Garcia PA,Wei T,Naym DG,Fredholm S,Fink-Puches R,Cerroni L,Odum N,O'Malley JT,Gniadecki R,Wolf P

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Immunohistochemistry

Fri Jul 01 00:00:00 EDT 2016

γ-Tocopherol supplementation of allergic female mice augments development of CD11c+CD11b+ dendritic cells in utero and allergic inflammation in neonates.

American journal of physiology. Lung cellular and molecular physiology

Abdala-Valencia H,Soveg F,Cook-Mills JM

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Fri Apr 15 00:00:00 EDT 2016

Group 2 innate lymphoid cells utilize the IRF4-IL-9 module to coordinate epithelial cell maintenance of lung homeostasis.

Mucosal immunology

Mohapatra A,Van Dyken SJ,Schneider C,Nussbaum JC,Liang HE,Locksley RM

14-9858 was used in Flow cytometry/Cell sorting to study how ILC2s link alveolar function to regulation of airway flow, revealing a key interaction between resident lymphoid and structural cells that might underlie similar organisational hierarchies in other organs.

Fri Jan 01 00:00:00 EST 2016

Co-potentiation of antigen recognition: A mechanism to boost weak T cell responses and provide immunotherapy in vivo.

Science advances

Hoffmann MM,Molina-Mendiola C,Nelson AD,Parks CA,Reyes EE,Hansen MJ,Rajagopalan G,Pease LR,Schrum AG,Gil D

14-9858 was used in Flow cytometry/Cell sorting to investigate co-potentiation, showing that weak T-cell responses can be boosted to provide immunotherapy in vivo.

Thu Oct 01 00:00:00 EDT 2015

Cell-intrinsic expression of TLR9 in autoreactive B cells constrains BCR/TLR7-dependent responses.

Journal of immunology (Baltimore, Md. : 1950)

Nündel K,Green NM,Shaffer AL,Moody KL,Busto P,Eilat D,Miyake K,Oropallo MA,Cancro MP,Marshak-Rothstein A

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Sun Mar 15 00:00:00 EDT 2015

Beta-catenin signaling drives differentiation and proinflammatory function of IRF8-dependent dendritic cells.

Journal of immunology (Baltimore, Md. : 1950)

Cohen SB,Smith NL,McDougal C,Pepper M,Shah S,Yap GS,Acha-Orbea H,Jiang A,Clausen BE,Rudd BD,Denkers EY

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Thu Jan 01 00:00:00 EST 2015

Comparative transcriptional and functional profiling defines conserved programs of intestinal DC differentiation in humans and mice.

Nature immunology

Watchmaker PB,Lahl K,Lee M,Baumjohann D,Morton J,Kim SJ,Zeng R,Dent A,Ansel KM,Diamond B,Hadeiba H,Butcher EC

14-9858 was used in Flow cytometry/Cell sorting to characterise human gut dendritic cell populations, and define their relationship to previously studied human and mouse dendritic cells.

Wed Jan 01 00:00:00 EST 2014

OX40 signaling favors the induction of T(H)9 cells and airway inflammation.

Nature immunology

Xiao X,Balasubramanian S,Liu W,Chu X,Wang H,Taparowsky EJ,Fu YX,Choi Y,Walsh MC,Li XC

14-9858 was used in Flow cytometry/Cell sorting to identify a previously unknown mechanism for the induction of Th9 cells, with potential clinical implications in allergic inflammation.

Mon Oct 01 00:00:00 EDT 2012

Regulatory T-cell suppressor program co-opts transcription factor IRF4 to control T(H)2 responses.

Nature

Zheng Y,Chaudhry A,Kas A,deRoos P,Kim JM,Chu TT,Corcoran L,Treuting P,Klein U,Rudensky AY

Published figure using IRF4 monoclonal antibody (Product # 14-9858-82) in Flow Cytometry

Thu Mar 19 00:00:00 EDT 2009

Bioinformatics

Protein Aliases: Interferon regulatory factor 4; IRF-4; LSIRF; Lymphocyte-specific interferon regulatory factor; Multiple myeloma oncogene 1; NF-EM5; PU.1 interaction partner; Sfpi1/PU.1 interaction partner; Transcriptional activator PIP

Gene Aliases: AI385587; IRF-4; IRF4; LSIRF; MUM1; NF-EM5; SHEP8; Spip

UniProt ID: (Human) Q15306, (Mouse) Q64287

Entrez Gene ID: (Human) 3662, (Mouse) 16364

Molecular Function: winged helix/forkhead transcription factor