IRF9 Recombinant Rabbit Monoclonal Antibody [PSH08-20]

☑ Cell treatment (CT)

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.

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.

☑ Cell treatment (CT)

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.

IRF9 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA722963 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722963 at 1/2,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) 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

☑ Cell treatment (CT)

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.