Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR27554-69]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR27554-69] (AB315277)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Interferon regulatory factor 9/IRF-9 with ab315277 at 1/1000 (0.48 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The section was incubated with ab315277 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR27554-69] (AB315277)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) treated with 10ng/ml IFN alpha for 24h (Red) / Untreated NIH/3T3 (Green) cells labelling Interferon regulatory factor 9/IRF-9 with ab315277 at 1/50 dilution (1 ug)/Red and green compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR27554-69] (AB315277)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Interferon regulatory factor 9/IRF-9 with ab315277 at 1/1000 (0.48 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse liver (PMID : 28442624). The section was incubated with ab315277 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR27554-69] (AB315277)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Interferon regulatory factor 9/IRF-9 with ab315277 at 1/50 (9.6 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/mL) dilution (Green).

Confocal image showing nuclear staining in NIH/3T3 cells treated with IFNα (mouse) (10 ng/mL) for 24 hr. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/mL) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR27554-69] (AB315277)

Immunohistochemical analysis of paraffin-embedded Mouse lung cancer tissue labeling Interferon regulatory factor 9/IRF-9 with ab315277 at 1/1000 (0.48 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung cancer. The section was incubated with ab315277 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

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Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR27554-69] (AB315277)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression of IRF-9 is upregulated in response to IFN alpha treatment.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR27554-69] (ab315277) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa treated with 10ng/ml IFN alpha for 16 hours whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 48 kDa,36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR27554-69] (AB315277)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the band at 37 kDa is unknown.

All lanes:

Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR27554-69] (ab315277) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 20 µg

Lane 3:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 48 kDa

true

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR27554-69] (AB315277)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression of IRF-9 is upregulated in response to IFN alpha treatment.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR27554-69] (ab315277) at 1/1000 dilution

Lane 1:

Untreated NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 2:

HeLa treated with 10ng/ml IFN alpha for 16 hours whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 48 kDa,36 kDa

false

Exposure time: 26s