WB
Western blot analysis of IRF3 on different lysates with Rabbit anti-IRF3 antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate (15 µg/Lane) Lane 2: Jurkat cell lysate (15 µg/Lane) Lane 3: MCF7 cell lysate (15 µg/Lane) Lane 4: A549 cell lysate (15 µg/Lane) Lane 5: THP-1 cell lysate (15 µg/Lane) Predicted band size: 47 kDa Observed band size: 55 kDa Exposure time: 43 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 1/5,000 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 mouse testis tissue with Rabbit anti-IRF3 antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 1/2,000 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 Jurkat cells labeling IRF3 with Rabbit anti-IRF3 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-IRF3 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 Jurkat cells labeling IRF3. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). 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℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
IRF3 was immunoprecipitated from 0.2 mg Jurkat cell lysate with at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Jurkat cell lysate (input) Lane 2:IRF3 antibody IP in Jurkat cell lysate Lane 3: Rabbit IgG instead of IRF3 antibody in Jurkat cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 9 seconds; ECL| Product Name | IRF3 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human IRF3 aa 71-120 / 427. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:1000-1:5000 IHC:1:50-1:2000 ICC:1:50-1:200 FC:1:1000 IP:1-2ug/sample |
|
| 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] |
| Alternative Names | IIAE7 antibody Interferon regulatory factor 3 antibody IRF 3 antibody IRF-3 antibody IRF3 antibody IRF3_HUMAN antibody MGC94729 antibody |
| Molecular Weight(MW) | 47kDa(Observed band size: 55kDa) |
| Cellular Localization | Cytoplasm, Nucleus. |
WB
Western blot analysis of IRF3 on different lysates with Rabbit anti-IRF3 antibody at 1/5,000 dilution. Lane 1: HeLa cell lysate (15 µg/Lane) Lane 2: Jurkat cell lysate (15 µg/Lane) Lane 3: MCF7 cell lysate (15 µg/Lane) Lane 4: A549 cell lysate (15 µg/Lane) Lane 5: THP-1 cell lysate (15 µg/Lane) Predicted band size: 47 kDa Observed band size: 55 kDa Exposure time: 43 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 1/5,000 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 mouse testis tissue with Rabbit anti-IRF3 antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 1/2,000 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 Jurkat cells labeling IRF3 with Rabbit anti-IRF3 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-IRF3 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 Jurkat cells labeling IRF3. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). 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℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
IRF3 was immunoprecipitated from 0.2 mg Jurkat cell lysate with at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Jurkat cell lysate (input) Lane 2:IRF3 antibody IP in Jurkat cell lysate Lane 3: Rabbit IgG instead of IRF3 antibody in Jurkat cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 9 seconds; ECL| Application Notes | WB:1:1000-1:5000 IHC:1:50-1:2000 ICC:1:50-1:200 FC:1:1000 IP:1-2ug/sample |
|---|
| 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. |
