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IRF4 Monoclonal Antibody (3E4), PE-eFluor™ 610, eBioscience™
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FIGURE: 1 / 17
IRF4 Antibody (61-9858-82) in Flow
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-... View More
Product Details
61-9858-82
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Human,
Mouse
Published species
Mouse,
Rhesus monkey
Host/Isotype
Rat
/ IgG1, kappa
Recommended Isotype Control
Class
Monoclonal
Type
Antibody
Clone
3E4
Conjugate
Excitation/Emission Max
565/606 nm
View spectra
Form
Liquid
Concentration
0.2 mg/mL
Amount
100 µg
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Ambient (domestic); Wet ice (international)
RRID
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), 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 (Product # 00-5523-00) and protocol. Please refer to BestProtocols®: 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 (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 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.
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.
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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; IMD131; IRF-4; IRF4; LSIRF; MUM1; NF-EM5; SHEP8; Spip
UniProt ID: (Human) Q15306, (Mouse) Q64287
Entrez Gene ID: (Human) 3662, (Mouse) 16364
transcription regulatory region sequence-specific DNA binding
RNA polymerase II core promoter proximal region sequence-specific DNA binding
RNA polymerase II transcription factor activity, sequence-specific DNA binding
core promoter proximal region sequence-specific DNA binding
transcriptional activator activity, RNA polymerase II transcription regulatory region sequence-specific binding
DNA binding
transcription factor activity, sequence-specific DNA binding
transcription coactivator activity
protein binding
sequence-specific DNA binding
sequence-specific double-stranded DNA binding
regulatory region DNA binding
transcriptional activator activity, RNA polymerase II core promoter proximal region sequence-specific binding
protein-lysine N-methyltransferase activity
winged helix/forkhead transcription factor
immune system process
chromatin remodeling
regulation of transcription, DNA-templated
regulation of transcription from RNA polymerase II promoter
positive regulation of interleukin-10 production
positive regulation of interleukin-13 production
positive regulation of interleukin-2 production
positive regulation of interleukin-4 production
negative regulation of toll-like receptor signaling pathway
T cell activation
defense response to protozoan
myeloid dendritic cell differentiation
regulation of T-helper cell differentiation
positive regulation of transcription, DNA-templated
positive regulation of transcription from RNA polymerase II promoter
positive regulation of multicellular organismal process
T-helper 17 cell lineage commitment
positive regulation of cold-induced thermogenesis
transcription, DNA-templated
transcription from RNA polymerase II promoter
protein methylation
peptidyl-lysine methylation
positive regulation of DNA binding
histone H3 acetylation
histone H4 acetylation
positive regulation of interleukin-10 biosynthetic process
positive regulation of interleukin-2 biosynthetic process
positive regulation of interleukin-13 biosynthetic process
positive regulation of interleukin-4 biosynthetic process
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