Antibody data
- Antibody Data
- Antigen structure
- References [6]
- Comments [0]
- Validations
- Flow cytometry [1]
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- Product number
- 61-9858-80 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-IRF4 Monoclonal Antibody (3E4), PE-eFluor 610, 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 Staining Buffer Set (cat. 00-5523) and protocol. Please refer to Best Protocols: Protocol B: One step protocol for (nuclear) intracellular proteins located under the Resources Tab online. This can be used at less than or equal to 0.5 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. PE-eFluor® 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red®). Please make sure that your instrument is capable of detecting this fluorochome. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green 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 ROCK2, but not ROCK1 interacts with phosphorylated STAT3 and co-occupies TH17/TFH gene promoters in TH17-activated human T cells.
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.
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.
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
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
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
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- Invitrogen Antibodies (provider)
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- Experimental details
- Normal human peripheral blood cells were stimulated for 3 days with Anti-Human CD3 and Anti-Human CD28 Functional Grade Purifieds (Product # 16-0037-81 and Product # 16-0289-81) and then intracellularly stained with 0.25 µg of Rat IgG1 K Isotype Control PE-eFluor® 610 (Product # 61-4301-82) (blue histogram) or 0.25 µg of Anti-Human/Mouse IRF4 PE-eFluor® 610 (purple histogram) using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. The orange histogram is unstimulated cells stained with 0.25 µg of Anti-Human/Mouse IRF4 PE-eFluor® 610. Viable lymphocytes, as determined by Fixable Viability Dye eFluor® 450 (Product # 65-0863-14), were used for analysis.