IRF9 Recombinant Rabbit Monoclonal Antibody [PSH08-20]
cat.: HA722963
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, IP, ChIP |
| Clonality: | Monoclonal |
| Clone number: | PSH08-20 |
| 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: 44 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human IRF9 aa 1-393. |
| Positive control: | HeLa cell lysate, HeLa treated with 10ng/mL IFN-α1 for 16 hours cell lysate, Daudi cell lysate, Daudi treated with 10ng/mL IFN-α1 for 16 hours cell lysate, Jurkat cell lysate, Jurkat treated with 100nM IFN-α1 for 24 hours cell lysate, THP-1 cell lysate, HeLa cells treated with 10ng/mL IFN-α1 for 16 hours. |
| Subcellular location: | Cytoplasm, Nucleus. |
| Recommended Dilutions:
WB IF-Cell IP ChIP |
1:1,000 1:100 1-2μg/sample Use 2~5 μg for 25 μg of chromatin. |
| Uniprot #: | SwissProt: Q00978 Human |
| Alternative names: | IFN alpha responsive transcription factor subunit IFN-alpha-responsive transcription factor subunit Interferon regulatory factor 9 interferon stimulated transcription factor 3 Interferon-stimulated gene factor 3 gamma interferon-stimulated transcription factor 3, gamma 48kDa IRF 9 IRF-9 Irf9 IRF9_HUMAN ISGF 3 gamma ISGF-3 gamma ISGF3 ISGF3 p48 subunit ISGF3G OTTHUMP00000164692 OTTHUMP00000164693 p48 Transcriptional regulator ISGF3 subunit gamma |
Images
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Fig1:
Western blot analysis of IRF9 on different lysates with Rabbit anti-IRF9 antibody (HA722963) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 10ng/mL IFN-α1 for 16 hours cell lysate Lane 3: Daudi cell lysate Lane 4: Daudi treated with 10ng/mL IFN-α1 for 16 hours cell lysate Lane 5: Jurkat cell lysate Lane 6: Jurkat treated with 100nM IFN-α1 for 24 hours cell lysate Lane 7: THP-1 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 44 kDa Observed band size: 48 kDa Exposure time: 46 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 (HA722963) 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. |
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Fig2:
Western blot analysis of IRF9 on different lysates with Rabbit anti-IRF9 antibody (HA722963) at 1/1,000 dilution. Lane 1: Human IRF9 recombinant protein, 30ng/Lane Lane 2: Human IRF8 recombinant protein, 30ng/Lane Exposure time: 5 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 (HA722963) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig3:
Immunocytochemistry analysis of HeLa cells treated with 10ng/mL IFN-α1 for 16 hours labeling IRF9 with Rabbit anti-IRF9 antibody (HA722963) 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 (HA722963) 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 (HA601187, 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. |
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Fig4:
IRF9 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA722963 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722963 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: HA722963 IP in HeLa cell lysate Lane 3: Rabbit IgG instead of HA722963 in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 1 minute; ECL: K1802 |
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Fig5: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 10ng/mL IFN-α1 for 16 hours with IRF9 (HA722963) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.