Phospho-IRF3 (S396) Recombinant Rabbit Monoclonal Antibody [PSH06-75]
cat.: HA722772
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH06-75 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 47 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser396 of human IRF3. |
| Positive control: | HT-29 transfected with 2.5μg/mL Poly(I:C) for 6 hours cell lysate, J774A.1 transfected with 2.5μg/mL Poly(I:C) for 6 hours cell lysate, human testis tissue, HT-29 cells transfected with 2.5μg/mL Poly(I:C) for 6 hours. |
| Subcellular location: | Cytoplasm, Nucleus, Mitochondrion. |
| Recommended Dilutions:
WB IHC-P IF-Cell FC |
1:1,000 1:200 1:100 1:1,000 |
| Uniprot #: | SwissProt: Q14653 Human | P70671 Mouse Entrez Gene: 292892 Rat |
| Alternative names: | IIAE7 Interferon regulatory factor 3 IRF 3 IRF-3 IRF3 IRF3_HUMAN MGC94729 |
Images
|
Fig1:
Western blot analysis of Phospho-IRF3 (S396) on different lysates with Rabbit anti-Phospho-IRF3 (S396) antibody (HA722772) 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. Predicted band size: 47 kDa Observed band size: 47-55 kDa Exposure time: 12 seconds; ECL: K1801; 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 (HA722772) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
|
Fig2:
Immunohistochemical analysis of paraffin-embedded human testis tissue untreated / treated with λpp with Rabbit anti-Phospho-IRF3 (S396) antibody (HA722772) 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 (HA722772) 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. |
|
Fig3:
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 (HA722772) 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 (HA722772) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
|
Fig4:
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 (HA722772, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.