WB
Western blot analysis of Phospho-IRF3 (S396) on different lysates with Rabbit anti-Phospho-IRF3 (S396) antibody at 1/1,000 dilution. Lane 1: HT-29 untransfected cell lysate, Lane 2: HT-29 transfected with 2.5μg/mL Poly (I:C) for 6 hours cell lysate, Lane 3: J774A.1 untransfected cell lysate, Lane 4: J774A.1 transfected with 2.5μg/mL Poly (I:C) for 6 hours cell lysate, Lane 5: HT-29 transfected with 2.5μg/mL Poly (I:C) for 6 hours cell lysate, then the membrane treated with λpp for 1 hour. Lysates/proteins at 20 µg/Lane. Exposure time: 12 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human testis tissue untreated / treated with λpp with Rabbit anti-Phospho-IRF3 (S396) antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of untreated HT-29 cells (top) / HT-29 cells transfected with 2.5μg/mL Poly (I:C) for 6 hours (bottom) labeling Phospho-IRF3 (S396) with Rabbit anti-Phospho-IRF3 (S396) antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-IRF3 (S396) antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of untreated HT-29 cells (top) / HT-29 cells transfected with 2.5μg/mL Poly (I:C) for 6 hours (bottom) labeling Phospho-IRF3 (S396). Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (right) compared with Rabbit IgG Isotype Control (left). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃.| Product Name | Phospho-IRF3 (S396) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic phospho-peptide corresponding to residues surrounding Ser396 of human IRF3. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:1000 IHC:1:200 ICC/IF:1:100 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | IRF3 |
|---|---|
| Gene Synonyms | IIAE7 |
| Gene Full Name | interferon regulatory factor 3 |
| Gene Summary | This gene encodes a member of the interferon regulatory transcription factor (IRF) family. The encoded protein is found in an inactive cytoplasmic form that upon serine/threonine phosphorylation forms a complex with CREBBP. This complex translocates to the nucleus and activates the transcription of interferons alpha and beta, as well as other interferon-induced genes. The protein plays an important role in the innate immune response against DNA and RNA viruses. Mutations in this gene are associated with Encephalopathy, acute, infection-induced, herpes-specific, 7. [provided by RefSeq, Sep 2020] |
| Molecular Weight(MW) | 47kDa(Observed band size: 47-55kDa) |
| Cellular Localization | Cytoplasm, Nucleus, Mitochondrion. |
WB
Western blot analysis of Phospho-IRF3 (S396) on different lysates with Rabbit anti-Phospho-IRF3 (S396) antibody at 1/1,000 dilution. Lane 1: HT-29 untransfected cell lysate, Lane 2: HT-29 transfected with 2.5μg/mL Poly (I:C) for 6 hours cell lysate, Lane 3: J774A.1 untransfected cell lysate, Lane 4: J774A.1 transfected with 2.5μg/mL Poly (I:C) for 6 hours cell lysate, Lane 5: HT-29 transfected with 2.5μg/mL Poly (I:C) for 6 hours cell lysate, then the membrane treated with λpp for 1 hour. Lysates/proteins at 20 µg/Lane. Exposure time: 12 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human testis tissue untreated / treated with λpp with Rabbit anti-Phospho-IRF3 (S396) antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of untreated HT-29 cells (top) / HT-29 cells transfected with 2.5μg/mL Poly (I:C) for 6 hours (bottom) labeling Phospho-IRF3 (S396) with Rabbit anti-Phospho-IRF3 (S396) antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-IRF3 (S396) antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of untreated HT-29 cells (top) / HT-29 cells transfected with 2.5μg/mL Poly (I:C) for 6 hours (bottom) labeling Phospho-IRF3 (S396). Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (right) compared with Rabbit IgG Isotype Control (left). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃.| Application Notes | WB:1:1000 IHC:1:200 ICC/IF:1:100 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
Data sheet for OM643866