Phospho-IRF3 (S386) and Total IRF3 ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-IRF3 (S386) and Total IRF3 in Human in Cell Lysate samples.
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Phospho-IRF3 (S386) and Total IRF3 ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-IRF3 (S386) and Total IRF3 in Human in Cell Lysate samples.
Phospho-IRF3 (S386) and Total IRF3 (S386) ELISA Kit (ab279834) is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated IRF3 protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-IRF3 (S386) and total IRF3. An anti-pan IRF3 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and IRF3 present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho IRF3 (S386) antibody is added to detect phosphorylated IRF3. In the remaining wells, mouse anti-pan-IRF3 antibody is used to detect pan IRF3. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-conjugated anti-mouse IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of IRF3 (S386) or pan IRF3 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
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IRF3 also known as Interferon Regulatory Factor 3 acts as an important transcription factor in the immune response. It has a molecular weight of approximately 47 kDa. The IRF3 protein is mainly expressed in the cytoplasm and nucleus of various cell types including immune cells such as macrophages and dendritic cells. The protein becomes activated through phosphorylation a process frequently identified in its phosphorylated form phospo-IRF3 or p-IRF3 which facilitates its role in immune function.
IRF3 participates in the regulation of type I interferon (IFN) response a fundamental antiviral defense mechanism. IRF3 when phosphorylated forms a complex with CBP/p300 which then translocates to the nucleus to drive the expression of IFN-stimulated genes. This action strengthens the innate immune response and boosts the body's ability to counteract viral infections. Its activity and regulation are significant for maintaining a balanced immune response without excessive inflammation.
IRF3 is involved in the Toll-like receptor (TLR) and RIG-I-like receptor (RLR) signaling pathways both essential in pathogen recognition and response. Within these pathways IRF3 interacts with proteins such as MAVS and TBK1 to propagate immune signaling. The activation of IRF3 in these pathways results in the production of type I interferons and other cytokines orchestrating an effective antiviral response. These interactions highlight the protein's central role in mediating immune signaling cascades.
IRF3's malfunction or deregulation can contribute to autoimmune diseases and antiviral deficiencies. Conditions such as systemic lupus erythematosus (SLE) and chronic hepatitis B infection are linked to IRF3 activity. During autoimmune responses or viral persistence the aberrant activation of IRF3 can lead to inappropriate immune responses. The connection with proteins like STAT1 in these conditions highlights the complex network IRF3 engages in facilitating its impact on disease progression and immune dysregulation.
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Positive Control.
HT29 cells were treated with Calyculin A.
Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer.
Serial dilutions of lysates were analyzed in this ELISA.
HT29 cells treated with Calyculin A. HT29 cells were treated or untreated with Calyculin A.
HT29 cells treated with Calyculin A. HT29 cells were treated or untreated with Calyculin A.
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