Confocal immunofluorescent analysis of HT-29 cells, mock transfected (left) or transfected with Poly(dA:dT) (2 μg/mL, 6 hr; right), using IRF-3 (D9J5Q) Mouse mAb (Alexa Fluor® 647 Conjugate) (red). Actin filaments were labeled with DyLight™ 488 Phalloidin #12935 (green). Blue = DAPI #4083 (fluorescent DNA dye). Note the nuclear translocation of IRF-3 in a subset of cells (arrows).
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct immunofluorescent analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated IRF-3 (D9J5Q) Mouse mAb #10949.
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.
NOTE: Formaldehyde is toxic, use only in a fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised November 2013
Protocol Id: 182
IRF-3 (D9J5Q) Mouse mAb (Alexa Fluor® 647 Conjugate) recognizes endogenous levels of total IRF-3 protein.
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to human IRF-3 protein.
Interferon regulatory factors (IRFs) comprise a family of transcription factors that function within the Jak/Stat pathway to regulate interferon (IFN) and IFN-inducible gene expression in response to viral infection (1). IRFs play an important role in pathogen defense, autoimmunity, lymphocyte development, cell growth, and susceptibility to transformation. The IRF family includes nine members: IRF-1, IRF-2, IRF-9/ISGF3γ, IRF-3, IRF-4 (Pip/LSIRF/ICSAT), IRF-5, IRF-6, IRF-7, and IRF-8/ICSBP. All IRF proteins share homology in their amino-terminal DNA-binding domains. IRF family members regulate transcription through interactions with proteins that share similar DNA-binding motifs, such as IFN-stimulated response elements (ISRE), IFN consensus sequences (ICS), and IFN regulatory elements (IRF-E) (2).
Explore pathways + proteins related to this product.