Antibody data
- Antibody Data
- Antigen structure
- References [10]
- Comments [0]
- Validations
- Flow cytometry [1]
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- Product number
- 50-9858-80 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-IRF4 Monoclonal Antibody (3E4), eFluor 660, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- 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), Th2, 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 intracellular staining followed by flow cytometric analysis. Applications Tested: This 3E4 antibody has been tested by intracellular staining and flow cytometric analysis of stimulated normal human peripheral blood cells using the Foxp3/Transcription Factor Buffer Set (cat 00-5523) and protocol. Please see Best Protocols Section (Staining intracellular Antigens for Flow Cytometry) for staining protocol (refer to Protocol B: One-step protocol for intracellular (nuclear) proteins. This can be used at less than or equal to 0.25 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. eFluor® 660 is a replacement for Alexa Fluor® 647. eFluor® 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human, Mouse
- Host
- Rat
- Isotype
- IgG
- Antibody clone number
- 3E4
- Vial size
- 25 µg
- Concentration
- 0.2 mg/mL
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references Pro-Survival Lipid Sphingosine-1-Phosphate Metabolically Programs T Cells to Limit Anti-tumor Activity.
Blimp-1 Functions as a Molecular Switch to Prevent Inflammatory Activity in Foxp3+RORγt+ Regulatory T Cells.
ROCK2, but not ROCK1 interacts with phosphorylated STAT3 and co-occupies TH17/TFH gene promoters in TH17-activated human T cells.
CD38-NAD+Axis Regulates Immunotherapeutic Anti-Tumor T Cell Response.
Batf3 selectively determines acquisition of CD8+ dendritic cell phenotype and function.
Affinity and dose of TCR engagement yield proportional enhancer and gene activity in CD4+ T cells.
STAT3/5-Dependent IL9 Overexpression Contributes to Neoplastic Cell Survival in Mycosis Fungoides.
γ-Tocopherol supplementation of allergic female mice augments development of CD11c+CD11b+ dendritic cells in utero and allergic inflammation in neonates.
Cell-intrinsic expression of TLR9 in autoreactive B cells constrains BCR/TLR7-dependent responses.
Beta-catenin signaling drives differentiation and proinflammatory function of IRF8-dependent dendritic cells.
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
Cell reports 2019 Aug 13;28(7):1879-1893.e7
Cell reports 2019 Aug 13;28(7):1879-1893.e7
Blimp-1 Functions as a Molecular Switch to Prevent Inflammatory Activity in Foxp3+RORγt+ Regulatory T Cells.
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
Cell reports 2018 Oct 2;25(1):19-28.e5
Cell reports 2018 Oct 2;25(1):19-28.e5
ROCK2, but not ROCK1 interacts with phosphorylated STAT3 and co-occupies TH17/TFH gene promoters in TH17-activated human T cells.
Chen W, Nyuydzefe MS, Weiss JM, Zhang J, Waksal SD, Zanin-Zhorov A
Scientific reports 2018 Nov 9;8(1):16636
Scientific reports 2018 Nov 9;8(1):16636
CD38-NAD+Axis Regulates Immunotherapeutic Anti-Tumor T Cell Response.
Chatterjee S, Daenthanasanmak A, Chakraborty P, Wyatt MW, Dhar P, Selvam SP, Fu J, Zhang J, Nguyen H, Kang I, Toth K, Al-Homrani M, Husain M, Beeson G, Ball L, Helke K, Husain S, Garrett-Mayer E, Hardiman G, Mehrotra M, Nishimura MI, Beeson CC, Bupp MG, Wu J, Ogretmen B, Paulos CM, Rathmell J, Yu XZ, Mehrotra S
Cell metabolism 2018 Jan 9;27(1):85-100.e8
Cell metabolism 2018 Jan 9;27(1):85-100.e8
Batf3 selectively determines acquisition of CD8+ dendritic cell phenotype and function.
Chandra J, Kuo PT, Hahn AM, Belz GT, Frazer IH
Immunology and cell biology 2017 Feb;95(2):215-223
Immunology and cell biology 2017 Feb;95(2):215-223
Affinity and dose of TCR engagement yield proportional enhancer and gene activity in CD4+ T cells.
Allison KA, Sajti E, Collier JG, Gosselin D, Troutman TD, Stone EL, Hedrick SM, Glass CK
eLife 2016 Jul 4;5
eLife 2016 Jul 4;5
STAT3/5-Dependent IL9 Overexpression Contributes to Neoplastic Cell Survival in Mycosis Fungoides.
Vieyra-Garcia PA, Wei T, Naym DG, Fredholm S, Fink-Puches R, Cerroni L, Odum N, O'Malley JT, Gniadecki R, Wolf P
Clinical cancer research : an official journal of the American Association for Cancer Research 2016 Jul 1;22(13):3328-39
Clinical cancer research : an official journal of the American Association for Cancer Research 2016 Jul 1;22(13):3328-39
γ-Tocopherol supplementation of allergic female mice augments development of CD11c+CD11b+ dendritic cells in utero and allergic inflammation in neonates.
Abdala-Valencia H, Soveg F, Cook-Mills JM
American journal of physiology. Lung cellular and molecular physiology 2016 Apr 15;310(8):L759-71
American journal of physiology. Lung cellular and molecular physiology 2016 Apr 15;310(8):L759-71
Cell-intrinsic expression of TLR9 in autoreactive B cells constrains BCR/TLR7-dependent responses.
Nündel K, Green NM, Shaffer AL, Moody KL, Busto P, Eilat D, Miyake K, Oropallo MA, Cancro MP, Marshak-Rothstein A
Journal of immunology (Baltimore, Md. : 1950) 2015 Mar 15;194(6):2504-12
Journal of immunology (Baltimore, Md. : 1950) 2015 Mar 15;194(6):2504-12
Beta-catenin signaling drives differentiation and proinflammatory function of IRF8-dependent dendritic cells.
Cohen SB, Smith NL, McDougal C, Pepper M, Shah S, Yap GS, Acha-Orbea H, Jiang A, Clausen BE, Rudd BD, Denkers EY
Journal of immunology (Baltimore, Md. : 1950) 2015 Jan 1;194(1):210-22
Journal of immunology (Baltimore, Md. : 1950) 2015 Jan 1;194(1):210-22
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Intracellular staining of 3-day Anti-Human CD3 and Anti-Human CD28 Functional Grade Purified (Product # 16-0031-82 and Product # 16-0281-82)-stimulated human peripheral blood cells with 0.125 µg of Rat IgG1 K Isotype Control eFluor® 660 (Product # 50-4301-82) (blue histogram) or 0.125 µg of Anti-Human/Mouse IRF4 eFluor® 660 (purple histogram). The orange histogram is unstimulated cells stained with 0.125 µg of Anti-Human/Mouse IRF4 eFluor® 660. Viable lymphocytes, as determined by Fixable Viability Dye eFluor® 450 (Product # 65-0863-14), were used for analysis.