WB
Western blot analysis of IRF9 on different lysates with Rabbit anti-IRF9 antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: HeLa treated with 10ng/mL IFN-α1 for 16 hours cell lysate (20 µg/Lane) Lane 3: Daudi cell lysate (20 µg/Lane) Lane 4: Daudi treated with 10ng/mL IFN-α1 for 16 hours cell lysate (20 µg/Lane) Lane 5: Jurkat cell lysate (20 µg/Lane) Lane 6: Jurkat treated with 100nM IFN-α1 for 24 hours cell lysate (20 µg/Lane) Lane 7: THP-1 cell lysate (20 µg/Lane) Lane 8: C2C12 cell lysate (20 µg/Lane) Lane 9: PC-12 cell lysate (20 µg/Lane) Lane 10: Mouse spleen tissue lysate (40 µg/Lane) Lane 11: Mouse lung tissue lysate (40 µg/Lane) Lane 12: Rat spleen tissue lysate (40 µg/Lane) Lane 13: Rat lung tissue lysate (40 µg/Lane) Predicted band size: 44 kDa Observed band size: 48 kDa Exposure time: 25 seconds; ECL 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 1/2,000 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 rat spleen tissue with Rabbit anti-IRF9 antibody at 1/200 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 1/200 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 HeLa cells treated with 10ng/mL IFN-α1 for 16 hours labeling IRF9 with Rabbit anti-IRF9 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-IRF9 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 HeLa cells treated with 10ng/mL IFN-α1 for 16 hours labeling IRF9. 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
IRF9 was immunoprecipitated from 0.2 mg HeLa cell lysate with at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: IRF9 antibody IP in HeLa cell lysate Lane 3: Rabbit IgG instead of IRF9 antibody in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 1 minute; ECL| Product Name | IRF9 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human IRF9 aa 1-393. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:2000 IHC:1:50-1:200 ICC:1:100 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | IRF9 |
|---|---|
| Gene Synonyms | p48 IRF-9 ISGF3 ISGF3G |
| Gene Full Name | interferon regulatory factor 9 |
| Gene Summary | This gene encodes a member of the interferon regulatory factor (IRF) family, a group of transcription factors with diverse roles, including virus-mediated activation of interferon, and modulation of cell growth, differentiation, apoptosis, and immune system activity. Members of the IRF family are characterized by a conserved N-terminal DNA-binding domain containing tryptophan (W) repeats. Mutations in this gene result in Immunodeficiency 65. [provided by RefSeq, Jul 2020] |
| Alternative Names | IFN alpha responsive transcription factor subunit antibody IFN-alpha-responsive transcription factor subunit antibody Interferon regulatory factor 9 antibody interferon stimulated transcription factor 3 antibody Interferon-stimulated gene factor 3 gamma antibody interferon-stimulated transcription factor 3 gamma 48kDa antibody IRF 9 antibody IRF-9 antibody Irf9 antibody IRF9_HUMAN antibody IFN alpha responsive transcription factor subunit antibody IFN-alpha-responsive transcription factor subunit antibody Interferon regulatory factor 9 antibody interferon stimulated transcription factor 3 antibody Interferon-stimulated gene factor 3 gamma antibody interferon-stimulated transcription factor 3 gamma 48kDa antibody IRF 9 antibody IRF-9 antibody Irf9 antibody IRF9_HUMAN antibody ISGF 3 gamma antibody ISGF-3 gamma antibody ISGF3 antibody ISGF3 p48 subunit antibody ISGF3G antibody OTTHUMP00000164692 antibody OTTHUMP00000164693 antibody p48 antibody Transcriptional regulator ISGF3 subunit gamma antibody |
| Molecular Weight(MW) | 44kDa(Observed band size: 48kDa) |
| Cellular Localization | Cytoplasm, Nucleus. |
WB
Western blot analysis of IRF9 on different lysates with Rabbit anti-IRF9 antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: HeLa treated with 10ng/mL IFN-α1 for 16 hours cell lysate (20 µg/Lane) Lane 3: Daudi cell lysate (20 µg/Lane) Lane 4: Daudi treated with 10ng/mL IFN-α1 for 16 hours cell lysate (20 µg/Lane) Lane 5: Jurkat cell lysate (20 µg/Lane) Lane 6: Jurkat treated with 100nM IFN-α1 for 24 hours cell lysate (20 µg/Lane) Lane 7: THP-1 cell lysate (20 µg/Lane) Lane 8: C2C12 cell lysate (20 µg/Lane) Lane 9: PC-12 cell lysate (20 µg/Lane) Lane 10: Mouse spleen tissue lysate (40 µg/Lane) Lane 11: Mouse lung tissue lysate (40 µg/Lane) Lane 12: Rat spleen tissue lysate (40 µg/Lane) Lane 13: Rat lung tissue lysate (40 µg/Lane) Predicted band size: 44 kDa Observed band size: 48 kDa Exposure time: 25 seconds; ECL 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 1/2,000 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 rat spleen tissue with Rabbit anti-IRF9 antibody at 1/200 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 1/200 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 HeLa cells treated with 10ng/mL IFN-α1 for 16 hours labeling IRF9 with Rabbit anti-IRF9 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-IRF9 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 HeLa cells treated with 10ng/mL IFN-α1 for 16 hours labeling IRF9. 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
IRF9 was immunoprecipitated from 0.2 mg HeLa cell lysate with at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: IRF9 antibody IP in HeLa cell lysate Lane 3: Rabbit IgG instead of IRF9 antibody in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 1 minute; ECL| Application Notes | WB:1:2000 IHC:1:50-1:200 ICC:1:100 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
