Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)

Rabbit Recombinant Monoclonal Interferon regulatory factor 9/IRF-9 antibody. Carrier free. Suitable for ICC/IF, IP, WB, IHC-P and reacts with Human samples. Cited in 1 publication.

Be the first to review this product! Submit a review

Images

Immunocytochemistry - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (AB282125), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	IP	WB	IHC-P	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested	Not recommended
Mouse	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Target data

See full target information IRF9

Function

Transcription factor that plays an essential role in anti-viral immunity. It mediates signaling by type I IFNs (IFN-alpha and IFN-beta). Following type I IFN binding to cell surface receptors, Jak kinases (TYK2 and JAK1) are activated, leading to tyrosine phosphorylation of STAT1 and STAT2. IRF9/ISGF3G associates with the phosphorylated STAT1:STAT2 dimer to form a complex termed ISGF3 transcription factor, that enters the nucleus. ISGF3 binds to the IFN stimulated response element (ISRE) to activate the transcription of interferon stimulated genes, which drive the cell in an antiviral state.

Alternative names

ISGF3G, IRF9, Interferon regulatory factor 9, IRF-9, IFN-alpha-responsive transcription factor subunit, ISGF3 p48 subunit, Interferon-stimulated gene factor 3 gamma, Transcriptional regulator ISGF3 subunit gamma, ISGF-3 gamma

Recommended products

Rabbit Recombinant Monoclonal Interferon regulatory factor 9/IRF-9 antibody. Carrier free. Suitable for ICC/IF, IP, WB, IHC-P and reacts with Human samples. Cited in 1 publication.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR24260-55
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C

Notes

ab282125 is the carrier-free version of Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Interferon regulatory factor 9 (IRF-9) also known as ISGF3-γ is a critical component in the transcriptional regulation of interferon-stimulated genes. This protein with a mass of approximately 48 kDa serves as an important mediator in the interferon signaling pathway. IRF-9 is widely expressed in various tissues where it functions as part of the immune response to pathogens. Its expression can be induced by interferons signaling molecules involved in antiviral defense.

Biological function summary

IRF-9 participates in the assembly of the transcription factor complex ISGF3 alongside STAT1 and STAT2. This complex translocates into the nucleus to initiate the transcription of interferon-stimulated genes that promote antiviral states. IRF-9 through this functional role impacts a range of cellular responses including proliferation apoptosis and immune regulation. It affects the temporal and spatial aspects of gene expression in response to interferons.

Pathways

IRF-9 plays a significant role in both the Jak-STAT and interferon signaling pathways. These pathways are essential for the activation of genes involved in immune defense. Through its involvement in these pathways IRF-9 interacts with proteins such as STAT1 and STAT2 creating a bridge between extracellular signals and gene activation. This involvement ensures that cells can respond effectively to viral infections enhancing innate and adaptive immune responses.

Associated diseases and disorders

IRF-9 has been implicated in autoimmune diseases and certain cancers. Dysfunction in the IRF-9 associated pathways can lead to conditions such as systemic lupus erythematosus where inefficient regulation of immune responses occurs. In cancer altered IRF-9 expression can contribute to uncontrolled cell proliferation. These connections highlight the importance of IRF-9 in maintaining cellular homeostasis and immune integrity. IRF-9's interaction with STAT proteins and interferon pathways highlights its central role in these disease processes.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

13 product images

Immunocytochemistry - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)
This data was developed using the same antibody clone in a different buffer formulation (Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043). Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 staining Interferon regulatory factor 9 in wild-type A549 cells and IRF9 knockout A549 cells treated with interferon-a1 (Recombinant human Interferon alpha 1 protein (Active) ab48750) at 10 ng/ml for 16 hours. The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 at 0.4 μg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)
False colour image of Western blot: Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 was shown to bind specifically to Interferon regulatory factor 9/IRF-9. A band was observed at 48 kDa in treated wild-type HeLa cell lysates with no signal observed at this size in IRF9 CRISPR-Cas9 edited cell line Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell line ab266051 (CRISPR-Cas9 edited cell lysate Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell lysate ab258472). The band observed in the CRISPR-Cas9 edited lysate lane below 48 kDa is likely to represent a truncated form of Interferon regulatory factor 9/IRF-9. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and IRF9 CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] (Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043) at 1/1000 dilution
Lane 1: wild-type HeLa Treated Interferon-alpha1 (hIFN-a1) (10 ng/ml, 16 h) cell lysate at 20 µg
Lane 2: IRF9 knockout HeLa treated: hIFN-a1 (10 ng/ml, 16 h) cell lysate at 20 µg
Lane 2: Western blot - Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell line (Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell line ab266051)
Lane 3: wild-type HeLa Control Interferon-alpha1 (hIFN-a1) (0 ng/ml, 16 h) cell lysate at 20 µg
Lane 4: IRF9 knockout HeLa vehicle control: hIFN-a1 (0 ng/ml, 16 h) cell lysate at 20 µg
Lane 5: THP-1 cell lysate at 20 µg
Lane 6: ACHN cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 44 kDa
Observed band size: 48 kDa
Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)
False colour image of Western blot: Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 was shown to bind specifically to Interferon regulatory factor 9/IRF-9. A band was observed at 48 kDa in treated wild-type A549 cell lysates with no signal observed at this size in IRF9 CRISPR-Cas9 edited cell line Human IRF9 (Interferon regulatory factor 9) knockout A549 cell line ab267119 (CRISPR-Cas9 edited cell lysate Human IRF9 (Interferon regulatory factor 9) knockout A549 cell lysate ab258473). The band observed in the CRISPR-Cas9 edited lysate lane below 48 kDa is likely to represent a truncated form of Interferon regulatory factor 9/IRF-9. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and IRF9 CRISPR-Cas9 edited A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] (Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043) at 1/1000 dilution
Lane 1: wild-type A549 Treated Interferon-alpha1 (hIFN-a1) (10 ng/ml, 16 h) cell lysate at 20 µg
Lane 2: IRF9 knockout A549 Treated Interferon-alpha1 (hIFN-a1) (10 ng/ml, 16 h) cell lysate at 20 µg
Lane 2: Western blot - Human IRF9 (Interferon regulatory factor 9) knockout A549 cell line (Human IRF9 (Interferon regulatory factor 9) knockout A549 cell line ab267119)
Lane 3: wild-type A549 Control Interferon-alpha1 (hIFN-a1) (0 ng/ml, 16 h) cell lysate at 20 µg
Lane 4: IRF9 knockout A549 Control Interferon-alpha1 (hIFN-a1) (0 ng/ml, 16 h) cell lysate at 20 µg
Lane 5: THP-1 cell lysate at 20 µg
Lane 6: ACHN cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 44 kDa
Observed band size: 48 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)
This data was developed using the same antibody clone in a different buffer formulation (Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043). Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 staining Interferon regulatory factor 9 in wild-type HeLa cells and IRF9 knockout HeLa cells treated with interferon-a1 (Recombinant human Interferon alpha 1 protein (Active) ab48750) at 10 ng/ml for 16 hours. The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 at 0.4 μg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)
This data was developed using Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
IRF9 expression is increased by IFN-alpha (PMID:18190617).
Exposure time: 15 seconds.
All lanes: Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] (Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043) at 1/1000 dilution
Lane 1: Untreated HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 40 µg
Lane 2: HeLa treated with 10ng/ml human IFN alpha 1 (Recombinant human Interferon alpha 1 protein (Active) ab48750) for 16 hours, whole cell lysate at 40 µg
Lane 3: THP-1 (human monocytic leukemia monocyte) whole cell lysate at 40 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 44 kDa
Observed band size: 48 kDa
Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)
This data was developed using Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
IFR9 can be degraded by the EV71-encoded 3C protease. A 48-kDa full length and 30-kDa cleaved IRF9 are observed. The molecular weight is consistent with what have been described in literature (PMID:21536800).
Exposure time: 3 minutes.
All lanes: Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] (Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043) at 1/1000 dilution
Lane 1: Human ovary cancer tissue lysate at 40 µg
Lane 2: Human liver cancer tissue lysate at 40 µg
Secondary
All lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution
Predicted band size: 44 kDa
Observed band size: 30 kDa, 48 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)
This data was developed using Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded HeLa (human cervix adenocarcinoma epithelial cell) cells (A) and HeLa cells treated by 10 ng/ml IFN for 18h (B) tissue labelling Interferon regulatory factor 9/IRF-9 with Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 at 1/2000 dilution (0.233 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Almost no staining on HeLa cells (A) and strongly positive staining on HeLa cells treated by 10 ng/ml IFN for 18h (B). The section was incubated with Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)
This data was developed using Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human spleen tissue labelling Interferon regulatory factor 9/IRF-9 with Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 at 1/2000 dilution (0.233 ug/ml) followed by a ready to use (Bond™ Polymer Refine Detection). Positive staining on human spleen. The section was incubated with Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunoprecipitation - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)
This data was developed using Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043, the same antibody clone in a different buffer formulation.
Interferon regulatory factor 9/IRF-9 was immunoprecipitated from 0.35 mg treated HeLa whole cell lysate with Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) treated with 10 ng/ml human IFN alpha 1 (Recombinant human Interferon alpha 1 protein (Active) ab48750) for 16 hours, whole cell lysate 10 ug
Lane 2: Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 IP in HeLa treated with 10 ng/ml human IFN alpha 1 (Recombinant human Interferon alpha 1 protein (Active) ab48750) for 16 hours whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 in HeLa treated with 10 ng/ml human IFN alpha 1 (Recombinant human Interferon alpha 1 protein (Active) ab48750) for 16 hours whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
All lanes: Immunoprecipitation - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] (Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043)
Predicted band size: 44 kDa
Observed band size: 48 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)
This data was developed using Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human lung tissue labelling Interferon regulatory factor 9/IRF-9 with Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 at 1/2000 dilution (0.233 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human lung (PMID: 30355451). The section was incubated with Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunoprecipitation - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)
This data was developed using Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043, the same antibody clone in a different buffer formulation.
Interferon regulatory factor 9/IRF-9 was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte) whole cell lysate with Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: THP-1 whole cell lysate 10 ug
Lane 2: Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 IP in THP-1 whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 in THP-1 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
All lanes: Immunoprecipitation - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] (Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043)
Predicted band size: 44 kDa
Observed band size: 48 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)
This data was developed using Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Clear cell carcinoma of human kidney tissue labelling Interferon regulatory factor 9/IRF-9 with Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 at 1/2000 dilution (0.233 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Clear cell carcinoma of human kidney (PMID: 30355451). The section was incubated with Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunocytochemistry/ Immunofluorescence - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] - BSA and Azide free (ab282125)
This data was developed using Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling Interferon regulatory factor 9/IRF-9 with Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] ab271043 at 1/50 dilution(9.3 ug/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/500 dilution (Green). Confocal image showing increased nuclear and weak cytoplasmic staining in HeLa cells treated with Interferon alpha-1 (10 ng/ml) for 16 h is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/500 dilution.